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<!--text--> | <!--text--> | ||
<div style="width:100%;padding:10% 30%;"> | <div style="width:100%;padding:10% 30%;"> | ||
− | <div style="width: | + | <div style="width:100%;"> |
<div> | <div> | ||
− | <p style="font-size: | + | <p style="font-size:48px; line-height:200%; margin:3pt 0pt; orphans:0; text-align:justify; widows:0"><span |
− | style="color:#1f497d; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#1f497d; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:48px; font-weight:bold">First Attempt to |
Construct the Plasmids</span></p> | Construct the Plasmids</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">A</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">milGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-weight:bold; "> |
PCR</span></p> | PCR</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0 ;text-align: center;"><img |
src="https://static.igem.org/mediawiki/2021/9/95/T--Shanghai_United--result01.jpg" width="605" height="422" alt="" | src="https://static.igem.org/mediawiki/2021/9/95/T--Shanghai_United--result01.jpg" width="605" height="422" alt="" | ||
style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | ||
</p> | </p> | ||
− | <p style="font-size: | + | <p style="font-size:16px; line-height:200%; margin:30px 0pt; orphans:0; text-align:center; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Figure 1: Gel |
Electrophoresis Results of PCR of </span><span | Electrophoresis Results of PCR of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold"> Genes</span></p> |
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Figure 1 shows gel |
electrophoresis results of </span><span | electrophoresis results of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PCR. Column M is a 2K </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">marker </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">ladder. Columns 1-6 are PCR |
products of </span><span | products of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">genes</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">All 1-6 columns displayed |
successful results at 700bp</span><span | successful results at 700bp</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> which </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">could be used for later |
experiments.</span><span | experiments.</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">ARS |
</span><span | </span><span | ||
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-weight:bold; ">Plasmids |
Extraction</span></p> | Extraction</span></p> | ||
<table cellspacing="0" cellpadding="0" style="border-collapse:collapse; margin:0pt auto;;text-align: center;"> | <table cellspacing="0" cellpadding="0" style="border-collapse:collapse; margin:0pt auto;;text-align: center;"> | ||
Line 275: | Line 275: | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-indent:21pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Group 1</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Group 2</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 292: | Line 292: | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSA</span> |
</p> | </p> | ||
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">425.9 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">307.8 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 312: | Line 312: | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSD</span> |
</p> | </p> | ||
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">293.4 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">300.8 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 332: | Line 332: | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSR</span> |
</p> | </p> | ||
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">214.4 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#999999; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#999999; border-left-style:solid; border-left-width:1pt; border-right-color:#999999; border-right-style:solid; border-right-width:1pt; border-top-color:#999999; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">154.0 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
− | <p style="font-size: | + | <p style="font-size:16px; line-height:200%; margin:30px 0pt; orphans:0; text-align:center; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Table 1: |
Concentrations of ARS Plasmids Extraction Results</span></p> | Concentrations of ARS Plasmids Extraction Results</span></p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Table 1 shows that all |
concentrations except the concentration of </span><span | concentrations except the concentration of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSR |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">from Group 2 are within |
the desired range. While Group 2 was collecting </span><span | the desired range. While Group 2 was collecting </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSR</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">, half of the sample was |
accidentally mixed with </span><span | accidentally mixed with </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSA |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">samples and must be |
− | discarded</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | discarded</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> - |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">hence the |
− | deviation.</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | deviation.</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">The concentrations were |
used in to calculate the amount of </span><span | used in to calculate the amount of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARS </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">genes to be added in restriction |
enzyme digestion. </span></p> | enzyme digestion. </span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">A</span><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">milGFP</span><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; "> |
</span><span | </span><span | ||
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-weight:bold; ">PCR |
Products Clean-up</span></p> | Products Clean-up</span></p> | ||
<table cellspacing="0" cellpadding="0" style="border-collapse:collapse; margin:0pt auto;text-align: center;"> | <table cellspacing="0" cellpadding="0" style="border-collapse:collapse; margin:0pt auto;text-align: center;"> | ||
Line 386: | Line 386: | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Group 1</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Group 2</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 403: | Line 403: | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span> |
</p> | </p> | ||
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">382.6 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">350 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
− | <p style="font-size: | + | <p style="font-size:16px; line-height:200%; margin:30px 0pt; orphans:0; text-align:center; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Table 2: |
Concentrations of </span><span | Concentrations of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold"> PCR Products |
Clean-up Results</span></p> | Clean-up Results</span></p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Table 2 shows the concentrations |
of products of </span><span | of products of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PCR clean-up. The concentrations of |
the samples are optimal and </span><span | the samples are optimal and </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">could </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">be used to calculate the |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">amount</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> of samples added when performing |
restriction enzyme digestion.</span></p> | restriction enzyme digestion.</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">ARS/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; "> |
</span><span | </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-weight:bold; ">Digestion</span> |
</p> | </p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0;text-align: center;"><img |
src="https://static.igem.org/mediawiki/2021/3/3a/T--Shanghai_United--result04.jpg" width="601" height="455" alt="" | src="https://static.igem.org/mediawiki/2021/3/3a/T--Shanghai_United--result04.jpg" width="601" height="455" alt="" | ||
style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | ||
</p> | </p> | ||
− | <p style="font-size: | + | <p style="font-size:16px; line-height:200%; margin:30px 0pt; orphans:0; text-align:center; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Figure 2: Gel |
− | Electrophoresis Results of Restriction Enzyme Digestion of | + | Electrophoresis Results of Restriction Enzyme Digestion of ARS Plasmids and amilGFP Genes</span><span |
− | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold"></span></p> | |
− | + | ||
− | + | ||
− | + | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | |
− | + | ||
− | + | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Figure 2 shows gel |
electrophoresis results of gel electrophoresis on restriction enzyme digestion products of </span><span | electrophoresis results of gel electrophoresis on restriction enzyme digestion products of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSA, ARSD, |
− | ARSR</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | ARSR</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">, and </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">genes. All products showed |
− | suc</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | suc</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">c</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">essful results</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> that could</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> be used for later |
− | experiments</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | experiments</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> and these |
− | target </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | target </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">genetic |
materials were extracted from the desired bands.</span><span | materials were extracted from the desired bands.</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">ARS/</span><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">amilGFP</span><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; "> |
</span><span | </span><span | ||
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-weight:bold; ">Gel |
Extraction</span></p> | Extraction</span></p> | ||
<table cellspacing="0" cellpadding="0" style="border-collapse:collapse;margin:0pt auto;text-align: center;"> | <table cellspacing="0" cellpadding="0" style="border-collapse:collapse;margin:0pt auto;text-align: center;"> | ||
Line 493: | Line 487: | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Group 1</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Group 2</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 510: | Line 504: | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSA</span> |
</p> | </p> | ||
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">31.0 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">28.1 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 530: | Line 524: | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSD</span> |
</p> | </p> | ||
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">48.3 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">23.3 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 550: | Line 544: | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSR</span> |
</p> | </p> | ||
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">5.2 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">20.5 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 570: | Line 564: | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:144.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span> |
</p> | </p> | ||
</td> | </td> | ||
<td colspan="2" | <td colspan="2" | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:295.5pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:top; width:295.5pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">106 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
Line 587: | Line 581: | ||
</tr> | </tr> | ||
</table> | </table> | ||
− | <p style="font-size: | + | <p style="font-size:16px; line-height:200%; margin:30px 0pt; orphans:0; text-align:center; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Table 3: the |
Concentrations of</span><span | Concentrations of</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">7.30 |
ARS/</span><span | ARS/</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Gel |
Extraction</span><span | Extraction</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">from</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Two Groups</span> |
</p> | </p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">We recycled</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">GFP </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">and</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">the three plasmids</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSA</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">,</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSD |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">and</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSR</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">after electrophoresis.</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">We used</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Nanodrop, a common lab |
spectrophotometer, to test the concentration of the four products.</span><span | spectrophotometer, to test the concentration of the four products.</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">According to Table 3, all data |
except the concentration of </span><span | except the concentration of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSR |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">from Group 1 are |
within</span><span | within</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">the</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">acceptable range.</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">A possible reason for the anomaly |
is that when the sample containing</span><span | is that when the sample containing</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSA |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">was</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">loaded into</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">the</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">gel box, the pipette</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">pierced</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">into the gel, causing</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">a small amount of |
leakage</span><span | leakage</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">of the sample.</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">It is noted that the concentration |
of </span><span | of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">is also abnormally high. The reason |
is that since the two samples of </span><span | is that since the two samples of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">used by the two groups are the |
same, we mixed them together and then tested </span><span | same, we mixed them together and then tested </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">theri</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">concentration.</span></p> |
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">After we found out the |
concentration of </span><span | concentration of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">,</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSA,</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSD</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">and</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSR</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">, it will be much easier to |
calculate the volume of </span><span | calculate the volume of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">needed to mix with each plasmid to |
construct T4 DNA ligase ligation system</span><span | construct T4 DNA ligase ligation system</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">according to the ratio between |
</span><span | </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">and</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">plasmid</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">is 1:10.</span></p> |
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#403ed6; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#403ed6; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-weight:bold; ">Colony |
PCR</span></p> | PCR</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0;text-align: center;"><img |
src="https://static.igem.org/mediawiki/2021/0/08/T--Shanghai_United--result06.jpg" width="605" height="286" alt="" | src="https://static.igem.org/mediawiki/2021/0/08/T--Shanghai_United--result06.jpg" width="605" height="286" alt="" | ||
style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | ||
</p> | </p> | ||
<p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | <p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Figure 3: Gel |
Electrophoresis Results of Colony PCR of First Experiment Attempt</span></p> | Electrophoresis Results of Colony PCR of First Experiment Attempt</span></p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">We</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">examined growths of |
ARS/</span><span | ARS/</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">, |
cultivated</span><span | cultivated</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">since</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Jul. 30. Results were suboptimal: |
only 4 colonies were found in a total of 6 plates. We then performed colony PCR and gel electrophoresis | only 4 colonies were found in a total of 6 plates. We then performed colony PCR and gel electrophoresis | ||
to test for ARS/</span><span | to test for ARS/</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> genes in observed</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">colonies. Figure 3 indicates that |
only one colony resulted to be ARSD/</span><span | only one colony resulted to be ARSD/</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> positive, indicating failure of |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">the </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">experimental procedure.</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:48px; line-height:200%; margin:3pt 0pt; orphans:0; text-align:justify; widows:0"><span |
− | style="color:#1f497d; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#1f497d; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:48px; font-weight:bold">Second Attempt to |
Construct the Plasmids</span></p> | Construct the Plasmids</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">A</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">milGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; "> |
</span><span | </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-weight:bold; ">Digestion</span> |
</p> | </p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><img |
src="https://static.igem.org/mediawiki/2021/7/7e/T--Shanghai_United--result07.jpg" width="464" height="271" alt="" | src="https://static.igem.org/mediawiki/2021/7/7e/T--Shanghai_United--result07.jpg" width="464" height="271" alt="" | ||
style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | ||
</p> | </p> | ||
<p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | <p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Figure 4: Gel |
− | Electrophoresis Results of | + | Electrophoresis Results of amilGFP</span><span |
− | + | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:20pt; font-style:italic; font-weight:bold"> | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:20pt; font-style:italic; font-weight:bold"> | ||
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">PCR in |
Attempt 2</span></p> | Attempt 2</span></p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">We performed PCR of</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> the</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">gene to prepare for </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">the </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">second retry of </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">the </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">experimental procedure, then we |
performed electrophoresis to test for </span><span | performed electrophoresis to test for </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PCR products.</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Figure 4 indicates that the second |
sample of group 1 resulted in failure, but the other samples are successful</span><span | sample of group 1 resulted in failure, but the other samples are successful</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> which could be retrieved for the |
later experiments use.</span></p> | later experiments use.</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">A</span><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; ">milGFP</span><span |
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold; "> |
</span><span | </span><span | ||
− | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-weight:bold; ">Gel |
Extraction</span></p> | Extraction</span></p> | ||
<table cellspacing="0" cellpadding="0" style="border-collapse:collapse;margin:0pt auto;text-align: center;"> | <table cellspacing="0" cellpadding="0" style="border-collapse:collapse;margin:0pt auto;text-align: center;"> | ||
Line 798: | Line 791: | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:219.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:219.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span> |
</p> | </p> | ||
</td> | </td> | ||
<td | <td | ||
style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:219.75pt"> | style="border-bottom-color:#000000; border-bottom-style:solid; border-bottom-width:1pt; border-left-color:#000000; border-left-style:solid; border-left-width:1pt; border-right-color:#000000; border-right-style:solid; border-right-width:1pt; border-top-color:#000000; border-top-style:solid; border-top-width:1pt; padding:5.5pt 2.5pt; vertical-align:middle; width:219.75pt"> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">92.3 ng/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">μL</span></p> |
</td> | </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
<p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | <p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Table 4: |
− | Concentrations of Gel Extraction of | + | Concentrations of Gel Extraction of amilGFP</span><span |
− | + | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:20pt; font-style:italic; font-weight:bold"> | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:20pt; font-style:italic; font-weight:bold"> | ||
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Enzyme |
Digestion Products</span></p> | Digestion Products</span></p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Table 4 showed the concentration |
of all products, extracted from gel from </span><span | of all products, extracted from gel from </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Digestion, combined. The result is |
− | optimal</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | optimal</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">and </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">could </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">be used to calculate the amount of |
sample added when performing T4 DNA Ligase Ligation. </span></p> | sample added when performing T4 DNA Ligase Ligation. </span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-weight:bold; ">Colony |
PCR</span></p> | PCR</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0;text-align: center;"><img |
src="https://static.igem.org/mediawiki/2021/3/3d/T--Shanghai_United--Proof_of_Concept04.jpg" width="605" height="458" alt="" | src="https://static.igem.org/mediawiki/2021/3/3d/T--Shanghai_United--Proof_of_Concept04.jpg" width="605" height="458" alt="" | ||
style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | ||
</p> | </p> | ||
<p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | <p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Figure 5: Gel |
Electrophoresis Results 1 of Colony PCR of Second Experimental Attempt</span></p> | Electrophoresis Results 1 of Colony PCR of Second Experimental Attempt</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0;text-align:center;"><img |
src="https://static.igem.org/mediawiki/2021/5/58/T--Shanghai_United--Proof_of_Concept05.jpg" width="605" height="458" alt="" | src="https://static.igem.org/mediawiki/2021/5/58/T--Shanghai_United--Proof_of_Concept05.jpg" width="605" height="458" alt="" | ||
style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | ||
</p> | </p> | ||
<p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | <p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Figure 6: Gel |
Electrophoresis Results 2 of Colony PCR of Second Experimental Attempt</span></p> | Electrophoresis Results 2 of Colony PCR of Second Experimental Attempt</span></p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Figures 5 and 6 show the result |
for colony PCR identification on the </span><span | for colony PCR identification on the </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E.coli</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> with </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARS/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> inserted that were cultivated |
previously. The purpose is to examine whether the </span><span | previously. The purpose is to examine whether the </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E.coli</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> contain</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">s</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">expected </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">gene segment of </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">ARSA/D/R</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> and </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">. </span></p> |
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-weight:normal">As showing |
above, a</span><span | above, a</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-weight:normal">ll samples are |
determined to have bands of corresponding molecular sizes, which shows that all selected colonies | determined to have bands of corresponding molecular sizes, which shows that all selected colonies | ||
yielded positive results of </span><span | yielded positive results of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic; font-weight:normal">ARS |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-weight:normal">and |
</span><span | </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic; font-weight:normal">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic; font-weight:normal"> |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-weight:normal">genes. |
</span></p> | </span></p> | ||
− | <p style="font-size: | + | <p style="font-size:48px; line-height:200%; margin:7.8pt 0pt 3pt; orphans:0; text-align:justify; widows:0"><span |
− | style="color:#1f497d; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#1f497d; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:48px; font-weight:bold">Function |
Tests</span></p> | Tests</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:30px 0pt; orphans:0; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">Function |
Test </span><span | Test </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">With</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; "> |
NaH</span><span | NaH</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">PO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">4</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">4</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">, |
Attempt 1</span></p> | Attempt 1</span></p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Such function tests were |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">perform</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">ed under the hypothesis that |
NaH</span><span | NaH</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> is </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">structurally </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">similar to arsenic compounds that |
were meant to be detected. NaH</span><span | were meant to be detected. NaH</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> is used to substitute for arsenic |
compounds due to experimental safety. No fluorescence was detected in all samples, indicating that the | compounds due to experimental safety. No fluorescence was detected in all samples, indicating that the | ||
concentrations of NaH</span><span | concentrations of NaH</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> may be too low or that genetically |
− | engin</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | engin</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">e</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">ered </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> will not react to |
NaH</span><span | NaH</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">.</span></p> |
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">Function |
Test </span><span | Test </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">With</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; "> |
NaH</span><span | NaH</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">PO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">4</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">4</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">, |
Attempt 2</span></p> | Attempt 2</span></p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">In order to verify our assumption |
in attempt 1, </span><span | in attempt 1, </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">NaH</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> was used again in this testing, |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">with greater concentration |
than the last test. No fluorescence was detected in all samples, indicating the inability of the | than the last test. No fluorescence was detected in all samples, indicating the inability of the | ||
genetically engineered </span><span | genetically engineered </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> to react with |
NaH</span><span | NaH</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">.</span></p> |
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span></p> |
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; widows:0"><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">Function |
Test </span><span | Test </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">With</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; "> |
C</span><span | C</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">H</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">6</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">6</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; ">AsNaO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">2</span> | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:15pt; ; vertical-align:sub">2</span> | ||
</p> | </p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; widows:0;text-align: center;"><img |
− | src="https://static.igem.org/mediawiki/2021/8/8e/T--Shanghai_United--Proof_of_Concept06.jpg" width=" | + | src="https://static.igem.org/mediawiki/2021/8/8e/T--Shanghai_United--Proof_of_Concept06.jpg" width="600" alt="" |
style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | style="-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | ||
</p> | </p> | ||
<p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | <p style="font-size:20pt; line-height:200%; margin:3pt 0pt; orphans:0; text-align:center; widows:0"><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">Figure |
</span><span | </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">7</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:16px; font-weight:bold">: Fluorescence |
Intensity of Transformed E. coli In Different Concentrations of C2H12AsNaO5 Solutions</span></p> | Intensity of Transformed E. coli In Different Concentrations of C2H12AsNaO5 Solutions</span></p> | ||
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Given that NaH</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">PO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> cannot be used to stimulate |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">the </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">expression of </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> in genetically engineered |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">, C</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">H</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">6</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">6</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">AsNaO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">5</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">5</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> was used for this function test. |
Extremely minor fluorescence was detected by a microplate reader after 19 hours of reaction time. This | Extremely minor fluorescence was detected by a microplate reader after 19 hours of reaction time. This | ||
result confirms the design of </span><span | result confirms the design of </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> to be correct and |
− | functional</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | functional</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">.</span></p> |
<p | <p | ||
− | style="font-size: | + | style="font-size:22px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
− | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Figure </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">7</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> displays the fluorescence |
intensity generated by a</span><span | intensity generated by a</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">n</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> ARSD/</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">amilGFP</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> transformed </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> reacting for 1 |
hour in C</span><span | hour in C</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">H</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">6</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">6</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">AsNaO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">5</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">5</span><span | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:11.5pt"> </span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:11.5pt"> </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">solutions. As seen in the figure, |
− | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">fluorescence</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> was detected for all C</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">H</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">6</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">6</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">AsNaO</span><span |
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">5</span><span | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">5</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> concentrations that are above 0, |
which confirms that the designed plasmid worked as intended. </span><span | which confirms that the designed plasmid worked as intended. </span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">However</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">the </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">fluorescence</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> was rather minor, </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">so we speculate </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">that </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> responds poorly |
to organic arsenic compounds. Further experiments</span><span | to organic arsenic compounds. Further experiments</span><span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">would </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">be</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> conducted </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">to test for </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">fluorescence</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> intensities of </span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> in</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> in</span><span |
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">organic arsenic solutions. </span> |
</p> | </p> | ||
<p | <p | ||
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<p | <p | ||
− | style="font-size: | + | style="font-size:36px; line-height:130%; margin:3pt 0pt; orphans:0; text-align:justify; text-indent:40pt; widows:0"> |
<span | <span | ||
− | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:36px; font-style:italic; font-weight:bold">To |
be continued ...</span></p> | be continued ...</span></p> | ||
<p | <p | ||
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− | <span style="font-size: 48px;display: block;padding-bottom: | + | <span |
+ | style="font-size: 48px;display: block;padding-bottom: 70px;">CONTACT INFO</span> | ||
<span | <span | ||
style="text-align: center;line-height: 90px;letter-spacing: 2px;font-size: 36px;display: block;font-family: -apple-system, BlinkMacSystemFont, 'Segoe UI', Roboto, Oxygen, Ubuntu, Cantarell, 'Open Sans', 'Helvetica Neue', sans-serif;"> | style="text-align: center;line-height: 90px;letter-spacing: 2px;font-size: 36px;display: block;font-family: -apple-system, BlinkMacSystemFont, 'Segoe UI', Roboto, Oxygen, Ubuntu, Cantarell, 'Open Sans', 'Helvetica Neue', sans-serif;"> | ||
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Weibo、Bilibili、Wechat ! | Weibo、Bilibili、Wechat ! | ||
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Revision as of 03:51, 6 October 2021
First Attempt to Construct the Plasmids
AmilGFP PCR
Figure 1: Gel Electrophoresis Results of PCR of amilGFP Genes
Figure 1 shows gel electrophoresis results of amilGFP PCR. Column M is a 2K marker ladder. Columns 1-6 are PCR products of amilGFP genes
All 1-6 columns displayed successful results at 700bp which could be used for later experiments.
ARS Plasmids Extraction
|
Group 1 |
Group 2 |
ARSA |
425.9 ng/μL |
307.8 ng/μL |
ARSD |
293.4 ng/μL |
300.8 ng/μL |
ARSR |
214.4 ng/μL |
154.0 ng/μL |
Table 1: Concentrations of ARS Plasmids Extraction Results
Table 1 shows that all concentrations except the concentration of ARSR from Group 2 are within the desired range. While Group 2 was collecting ARSR, half of the sample was accidentally mixed with ARSA samples and must be discarded - hence the deviation. The concentrations were used in to calculate the amount of ARS genes to be added in restriction enzyme digestion.
AmilGFP PCR Products Clean-up
|
Group 1 |
Group 2 |
amilGFP |
382.6 ng/μL |
350 ng/μL |
Table 2: Concentrations of amilGFP PCR Products Clean-up Results
Table 2 shows the concentrations of products of amilGFP PCR clean-up. The concentrations of the samples are optimal and could be used to calculate the amount of samples added when performing restriction enzyme digestion.
ARS/amilGFP Digestion
Figure 2: Gel Electrophoresis Results of Restriction Enzyme Digestion of ARS Plasmids and amilGFP Genes
Figure 2 shows gel electrophoresis results of gel electrophoresis on restriction enzyme digestion products of ARSA, ARSD, ARSR, and amilGFP genes. All products showed successful results that could be used for later experiments and these target genetic materials were extracted from the desired bands.
ARS/amilGFP Gel Extraction
|
Group 1 |
Group 2 |
ARSA |
31.0 ng/μL |
28.1 ng/μL |
ARSD |
48.3 ng/μL |
23.3 ng/μL |
ARSR |
5.2 ng/μL |
20.5 ng/μL |
amilGFP |
106 ng/μL |
|
Table 3: the Concentrations of 7.30 ARS/amilGFP Gel Extraction from Two Groups
We recycled GFP and the three plasmids ARSA, ARSD and ARSR after electrophoresis. We used Nanodrop, a common lab spectrophotometer, to test the concentration of the four products. According to Table 3, all data except the concentration of ARSR from Group 1 are within the acceptable range. A possible reason for the anomaly is that when the sample containing ARSA was loaded into the gel box, the pipette pierced into the gel, causing a small amount of leakage of the sample. It is noted that the concentration of amilGFP is also abnormally high. The reason is that since the two samples of amilGFP used by the two groups are the same, we mixed them together and then tested theri concentration.
After we found out the concentration of amilGFP, ARSA, ARSD and ARSR, it will be much easier to calculate the volume of amilGFP needed to mix with each plasmid to construct T4 DNA ligase ligation system according to the ratio between amilGFP and plasmid is 1:10.
Colony PCR
Figure 3: Gel Electrophoresis Results of Colony PCR of First Experiment Attempt
We examined growths of ARS/amilGFP E. coli, cultivated since Jul. 30. Results were suboptimal: only 4 colonies were found in a total of 6 plates. We then performed colony PCR and gel electrophoresis to test for ARS/amilGFP genes in observed E. coli colonies. Figure 3 indicates that only one colony resulted to be ARSD/amilGFP positive, indicating failure of the experimental procedure.
Second Attempt to Construct the Plasmids
AmilGFP Digestion
Figure 4: Gel Electrophoresis Results of amilGFP PCR in Attempt 2
We performed PCR of the amilGFP gene to prepare for the second retry of the experimental procedure, then we performed electrophoresis to test for amilGFP PCR products. Figure 4 indicates that the second sample of group 1 resulted in failure, but the other samples are successful which could be retrieved for the later experiments use.
AmilGFP Gel Extraction
amilGFP |
92.3 ng/μL |
Table 4: Concentrations of Gel Extraction of amilGFP Enzyme Digestion Products
Table 4 showed the concentration of all products, extracted from gel from amilGFP Digestion, combined. The result is optimal and could be used to calculate the amount of sample added when performing T4 DNA Ligase Ligation.
Colony PCR
Figure 5: Gel Electrophoresis Results 1 of Colony PCR of Second Experimental Attempt
Figure 6: Gel Electrophoresis Results 2 of Colony PCR of Second Experimental Attempt
Figures 5 and 6 show the result for colony PCR identification on the E.coli with ARS/amilGFP inserted that were cultivated previously. The purpose is to examine whether the E.coli contains expected gene segment of ARSA/D/R and amilGFP.
As showing above, all samples are determined to have bands of corresponding molecular sizes, which shows that all selected colonies yielded positive results of ARS and amilGFP genes.
Function Tests
Function Test With NaH2PO4, Attempt 1
Such function tests were performed under the hypothesis that NaH2PO4 is structurally similar to arsenic compounds that were meant to be detected. NaH2PO4 is used to substitute for arsenic compounds due to experimental safety. No fluorescence was detected in all samples, indicating that the concentrations of NaH2PO4 may be too low or that genetically engineered E. coli will not react to NaH2PO4.
Function Test With NaH2PO4, Attempt 2
In order to verify our assumption in attempt 1, NaH2PO4 was used again in this testing, with greater concentration than the last test. No fluorescence was detected in all samples, indicating the inability of the genetically engineered E. coli to react with NaH2PO4.
Function Test With C2H6AsNaO2
Figure 7: Fluorescence Intensity of Transformed E. coli In Different Concentrations of C2H12AsNaO5 Solutions
Given that NaH2PO4 cannot be used to stimulate the expression of amilGFP in genetically engineered E. coli, C2H6AsNaO5 was used for this function test. Extremely minor fluorescence was detected by a microplate reader after 19 hours of reaction time. This result confirms the design of E. coli to be correct and functional.
Figure 7 displays the fluorescence intensity generated by an ARSD/amilGFP transformed E. coli reacting for 1 hour in C2H6AsNaO5 solutions. As seen in the figure, fluorescence was detected for all C2H6AsNaO5 concentrations that are above 0, which confirms that the designed plasmid worked as intended. However the fluorescence was rather minor, so we speculate that E. coli responds poorly to organic arsenic compounds. Further experiments would be conducted to test for fluorescence intensities of E. coli in inorganic arsenic solutions.
To be continued ...