Line 97: | Line 97: | ||
height: 1080px !important; | height: 1080px !important; | ||
} | } | ||
+ | ol li{ | ||
+ | list-style: decimal !important; | ||
+ | } | ||
+ | |||
</style> | </style> | ||
Line 223: | Line 227: | ||
<!--text--> | <!--text--> | ||
<div style="width:100%;padding:10% 30%;"> | <div style="width:100%;padding:10% 30%;"> | ||
− | <p style="font-size: | + | <p style="font-size:48px; margin: 0; orphans:0; text-align:center; 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:48px; font-weight:bold">Notebook</span></p> |
</hr> | </hr> | ||
− | <div style="width: | + | <div style="width:100%;margin: 100pt auto;"> |
<div> | <div> | ||
<p style="margin:0pt; orphans:0; text-align:center; widows:0"><img | <p style="margin:0pt; orphans:0; text-align:center; widows:0"><img | ||
src="https://static.igem.org/mediawiki/2021/0/07/T--Shanghai_United--Notebook01.jpg" width="70%" | src="https://static.igem.org/mediawiki/2021/0/07/T--Shanghai_United--Notebook01.jpg" width="70%" | ||
alt="c6e6151cedecf75dbdbc9b3e079800c" | alt="c6e6151cedecf75dbdbc9b3e079800c" | ||
− | style="width: 600px;-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | + | style="width: 600px;margin-bottom: 60px;-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:36px; line-height:130%; margin:40px 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-weight:bold;">Jul. |
28, 2021</span></p> | 28, 2021</span></p> | ||
− | <ol type="1" style="margin:0pt; padding-left:0pt"> | + | <ol type="1" start="1" style="text-align:justify;margin:0pt; padding-left:0pt"> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Learnt and clarified research goals and |
methods as a team. Learnt safety precautions necessary for entering the laboratory. </span></li> | methods as a team. Learnt safety precautions necessary for entering the laboratory. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Learnt and practiced the use of |
micropipettes. </span></li> | micropipettes. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed PCR of amilGFP gene; performed gel |
electrophoresis of said PCR products and tested for results. </span></li> | electrophoresis of said PCR products and tested for results. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Researched and learnt the reaction |
mechanisms of PCR and the preparation protocol of LB cultivation medium. </span></li> | mechanisms of PCR and the preparation protocol of LB cultivation medium. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Used LB cultivation medium with KANA |
antibiotics to cultivate three </span><span | antibiotics to cultivate three </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:22px; font-style:italic">E. coli</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:22px"> cell lines that are KANA resistant and with |
ArsA, ArsD, and ArsR genes inserted, respectively. </span></li> | ArsA, ArsD, and ArsR genes inserted, respectively. </span></li> | ||
</ol> | </ol> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:40px 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-weight:bold;">Jul. |
29, 2021</span></p> | 29, 2021</span></p> | ||
− | <ol type="1" style="margin:0pt; padding-left:0pt"> | + | <ol type="1" style="text-align:justify;margin:0pt; padding-left:0pt"> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed plasmid extraction of three |
− | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> cell lines, cultivated |
− | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">since </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:22px">Jul. 28. Said bacteria have KANA resistance |
and ArsA, ArsD, or ArsR depending on the cell line. Extracted plasmids are tested for and yielded | and ArsA, ArsD, or ArsR depending on the cell line. Extracted plasmids are tested for and yielded | ||
desirable concentrations. </span></li> | desirable concentrations. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Cleaned up amilGFP PCR products, |
− | obtained</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | obtained</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> in</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:22px"> Jul. 28. Cleaned up PCR products are tested |
for and yielded desirable concentrations. </span></li> | for and yielded desirable concentrations. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Using restriction enzyme BsaI, digested |
ArsA, ArsD, and ArsR plasmids. Reaction mix is left overnight to react. </span></li> | ArsA, ArsD, and ArsR plasmids. Reaction mix is left overnight to react. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Using restriction enzymes BsaI and DpnI, |
digested cleaned up amilGFP PCR products. Reaction mix is left overnight to react. </span></li> | digested cleaned up amilGFP PCR products. Reaction mix is left overnight to react. </span></li> | ||
</ol> | </ol> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:40px 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-weight:bold;">Jul. |
30, 2021</span></p> | 30, 2021</span></p> | ||
− | <ol type="1" style="margin:0pt; padding-left:0pt"> | + | <ol type="1" style="text-align:justify;margin:0pt; padding-left:0pt"> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Harvested restriction enzyme digestion |
products of ArsA, ArsD, and ArsR plasmids as well as amilGFP PCR products; digestion was | products of ArsA, ArsD, and ArsR plasmids as well as amilGFP PCR products; digestion was | ||
− | conducted</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | conducted</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> in</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:22px"> Jul. 29. </span></li> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed gel electrophoresis on restriction |
enzyme digestion products of ArsA, ArsD, ArsR, and amilGFP genes. </span></li> | enzyme digestion products of ArsA, ArsD, ArsR, and amilGFP genes. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed agarose gel DNA extraction of gel |
containing restriction enzyme digestion products. Extracted ArsA, ArsD, ArsR, and amilGFP genes are | containing restriction enzyme digestion products. Extracted ArsA, ArsD, ArsR, and amilGFP genes are | ||
tested for and yielded suboptimal concentrations. </span></li> | tested for and yielded suboptimal concentrations. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed T4 DNA ligase ligation of Ars |
genes and amilGFP genes. </span></li> | genes and amilGFP genes. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Using ligation products of Ars genes and |
amilGFP genes, transformed DH5-α </span><span | amilGFP genes, transformed DH5-α </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:22px; font-style:italic">E. coli</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:22px"> by heat shock method. </span></li> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Prepared LB cultivation medium agar plates |
with 0.1% KANA antibiotics. </span></li> | with 0.1% KANA antibiotics. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Spread Ars/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:22px; font-style:italic">E. coli</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:22px"> onto KANA antibiotics agar plates. </span> |
</li> | </li> | ||
</ol> | </ol> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:40px 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-weight:bold;">Jul. |
31, 2021</span></p> | 31, 2021</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt auto; orphans:0; text-align:center; widows:0"><img |
src="https://static.igem.org/mediawiki/2021/0/08/T--Shanghai_United--Notebook02.jpg" width="70%" alt="" | src="https://static.igem.org/mediawiki/2021/0/08/T--Shanghai_United--Notebook02.jpg" width="70%" alt="" | ||
style="width: 600px;-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | style="width: 600px;-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | ||
Line 339: | Line 343: | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:14pt; font-weight:bold">Gel | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:14pt; font-weight:bold">Gel | ||
Electrophoresis Results of amilGFP PCR</span></p> | Electrophoresis Results of amilGFP PCR</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:30pt auto; orphans:0; text-align:center; widows:0"><img |
src="https://static.igem.org/mediawiki/2021/7/76/T--Shanghai_United--Notebook03.jpg" width="70%" alt="" | src="https://static.igem.org/mediawiki/2021/7/76/T--Shanghai_United--Notebook03.jpg" width="70%" alt="" | ||
style="width: 600px;-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | style="width: 600px;-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | ||
Line 346: | Line 350: | ||
style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:14pt; font-weight:bold">Gel | style="color:#333333; font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:14pt; font-weight:bold">Gel | ||
Electrophoresis Results of Colony PCR</span></p> | Electrophoresis Results of Colony PCR</span></p> | ||
− | <ol type="1" style="margin:0pt; padding-left:0pt"> | + | <ol type="1" style="text-align:justify;margin:0pt; padding-left:0pt"> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Examined growths of Ars/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:22px; font-style:italic">E. coli</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:22px">, cultivated </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:22px">since </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:22px">Jul. 30. Results were suboptimal: only 4 |
colonies were found in a total of 6 plates.</span></li> | colonies were found in a total of 6 plates.</span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed colony PCR and gel electrophoresis |
to test for Ars/amilGFP genes in observed </span><span | to test for Ars/amilGFP genes in observed </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:22px; font-style:italic">E. coli</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:22px"> colonies. Only one colony resulted to be |
ArsD/amilGFP positive, indicating failure of experimental procedure. </span></li> | ArsD/amilGFP positive, indicating failure of experimental procedure. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed amilGFP restriction enzyme |
digestion on cleaned up amilGFP PCR products, obtained</span><span | digestion on cleaned up amilGFP PCR products, obtained</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:22px"> in</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:22px"> Jul. 29, to prepare for first retry of |
experimental procedure. </span></li> | experimental procedure. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed PCR of amilGFP gene to prepare for |
second retry of experimental procedure. Performed electrophoresis to test for amilGFP PCR products. | second retry of experimental procedure. Performed electrophoresis to test for amilGFP PCR products. | ||
</span></li> | </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed gel electrophoresis to test for |
amilGFP restriction enzyme digestion products. Performed agarose gel DNA extraction for digested | amilGFP restriction enzyme digestion products. Performed agarose gel DNA extraction for digested | ||
amilGFP genes. </span></li> | amilGFP genes. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Cleaned up amilGFP PCR products. </span> |
</li> | </li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed T4 DNA ligase ligation of digested |
amilGFP genes and digested Ars genes, obtained </span><span | amilGFP genes and digested Ars genes, obtained </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:22px">in </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:22px">Jul. 30. </span></li> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Cultivated three cell lines of </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:22px; font-style:italic">E. coli</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:22px"> with KANA resistance and ArsA, ArsD, or ArsR |
genes inserted, depending on the cell line, to prepare for second retry of experimental procedure. | genes inserted, depending on the cell line, to prepare for second retry of experimental procedure. | ||
</span></li> | </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Using ligation products of Ars genes and |
amilGFP genes, transformed DH5-α E. coli by heat shock method. </span></li> | amilGFP genes, transformed DH5-α E. coli by heat shock method. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 19.2pt; orphans:0; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Spread Ars/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:22px; font-style:italic">E. coli</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:22px"> onto KANA antibiotics agar plates, prepared |
− | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">in </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:22px">Jul. 30. </span></li> |
</ol> | </ol> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:40px 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-weight:bold;">Aug. |
1, 2021</span></p> | 1, 2021</span></p> | ||
− | <ol type="1" style="margin:0pt; padding-left:0pt"> | + | <ol type="1" style="text-align:justify;margin:0pt; padding-left:0pt"> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Examined growths of Ars/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:22px; font-style:italic">E. coli</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:22px">, cultivated </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:22px">since </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:22px">Jul. 3</span><a style="color:#000000" |
− | href="h"><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | href="h"><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">1</span></a><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">. Results were optima</span><a |
− | style="color:#000000" href="h"><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | style="color:#000000" href="h"><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">l: |
colonies were observed on all six plates</span></a><span | colonies were observed on all six plates</span></a><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></li> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Performed colony PCR and gel electrophoresis |
to test for Ars/amilGFP genes in observed </span><span | to test for Ars/amilGFP genes in observed </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:22px; font-style:italic">E. coli</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:22px"> colonies. All selected colonies yielded |
positive results of Ars genes. </span></li> | positive results of Ars genes. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Extracted ArsA, ArsD, and ArsR plasmids from |
− | Ars </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | Ars </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">, cultivated Jul. 31. Because |
the first retry of experimental procedure yielded positive results, the second retry had been | the first retry of experimental procedure yielded positive results, the second retry had been | ||
terminated. </span></li> | terminated. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Cultivated Ars/amilGFP positive </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:22px; font-style:italic">E. coli</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:22px">. </span></li> |
</ol> | </ol> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:40px 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-weight:bold;">Aug. |
2, 2021</span></p> | 2, 2021</span></p> | ||
− | <ol type="1" style="margin:0pt; padding-left:0pt"> | + | <ol type="1" style="text-align:justify;margin:0pt; padding-left:0pt"> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Preserved Ars/amilGFP positive </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:22px; font-style:italic">E. coli</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:22px"> for future studies. </span></li> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Conducted function test of Ars/amilGFP |
− | positive </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | positive </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> with different |
concentrations of NaH</span><span | concentrations of NaH</span><span | ||
style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</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:22px">PO</span><span |
style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</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:22px"> solutions. </span></li> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Mailed Ars/amilGFP positive </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:22px; font-style:italic">E. coli</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:22px"> samples to sequencing companies to perform |
Sanger sequencing of Ars/amilGFP genes. </span></li> | Sanger sequencing of Ars/amilGFP genes. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Started construction of team Wikipedia |
reports. </span></li> | reports. </span></li> | ||
</ol> | </ol> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:40px 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-weight:bold;">Aug. |
3, 2021</span></p> | 3, 2021</span></p> | ||
− | <ol type="1" style="margin:0pt; padding-left:0pt"> | + | <ol type="1" style="text-align:justify;margin:0pt; padding-left:0pt"> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Examined function test results, conducted |
− | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">in </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:22px">Aug. 2. No green fluorescence was found in all |
tubes. </span></li> | tubes. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Conducted second function test of |
Ars/amilGFP positive </span><span | Ars/amilGFP positive </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:22px; font-style:italic">E. coli</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:22px"> with higher concentrations of NaH</span><span |
style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</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:22px">PO</span><span |
style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</span><span | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">4</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:22px">. </span></li> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Continued construction of team Wikipedia. |
</span></li> | </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Created product design information and user |
manual sheet for dry team for reference. </span></li> | manual sheet for dry team for reference. </span></li> | ||
</ol> | </ol> | ||
− | <p style="font-size: | + | <p style="font-size:36px; line-height:130%; margin:40px 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-weight:bold;">Aug. |
4, 2021</span></p> | 4, 2021</span></p> | ||
− | <p style="font-size: | + | <p style="font-size:22px; line-height:130%; margin:3pt auto; orphans:0; text-align:center; widows:0"><img |
src="https://static.igem.org/mediawiki/2021/f/fe/T--Shanghai_United--Notebook04.jpg" width="70%" | src="https://static.igem.org/mediawiki/2021/f/fe/T--Shanghai_United--Notebook04.jpg" width="70%" | ||
alt="d3ee87f2c64666dd414a0897fc4172d" | alt="d3ee87f2c64666dd414a0897fc4172d" | ||
− | style="width: 600px;-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> | + | style="width: 600px;margin-bottom: 50px;-aw-left-pos:0pt; -aw-rel-hpos:column; -aw-rel-vpos:paragraph; -aw-top-pos:0pt; -aw-wrap-type:inline" /> |
</p> | </p> | ||
− | <ol type="1" style="margin:0pt; padding-left:0pt"> | + | <ol type="1" style="text-align:justify;margin:0pt; padding-left:0pt"> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Examined second function test results, |
− | conducted</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | conducted</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> in</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:22px"> Aug. 3. No green fluorescence was found in |
all tubes, indicating the inability of Ars/amilGFP </span><span | all tubes, indicating the inability of Ars/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:22px; font-style:italic">E. coli</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:22px"> to detect phosphate. </span></li> |
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Analyzed results of Ars/amilGFP gene |
− | sequencing results, sequenced </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | sequencing results, sequenced </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">in |
− | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Aug. 2. Constructed Ars/amilGFP genes |
are correct. </span></li> | are correct. </span></li> | ||
<li | <li | ||
− | 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; line-height:130%; margin:3pt 0pt 3pt 16.4pt; orphans:0; padding-left:2.8pt; text-indent:0pt; widows:0"> |
− | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | <span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px">Conducted third function test of Ars/amilGFP |
− | positive </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | positive </span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px; font-style:italic">E. |
− | coli</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size: | + | coli</span><span style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:22px"> with different |
concentrations of C</span><span | concentrations of C</span><span | ||
style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</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:22px">H</span><span |
style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">6</span><span | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">6</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:22px">AsNaO</span><span |
style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</span><span | style="font-family:'Segoe UI', Tahoma, Geneva, Verdana, sans-serif; font-size:7.67pt; vertical-align:sub">2</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:22px">.</span></li> |
</ol> | </ol> | ||
</div> | </div> | ||
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<div style="background-color: #F9F9F9;"> | <div style="background-color: #F9F9F9;"> | ||
<div style="width: 80%;height: 1080px;margin: 0 auto;padding: 10% 0;"> | <div style="width: 80%;height: 1080px;margin: 0 auto;padding: 10% 0;"> | ||
− | <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 ! | ||
</span> | </span> | ||
− | <img style="width:810px;height: 345px;margin: 100px auto;display: block;" | + | <img style="width:810px;height: 345px;margin: 100px auto;display: block;" src="https://static.igem.org/mediawiki/2021/1/1d/T--Shanghai_United--homepage06.jpg"> |
− | + | ||
</div> | </div> | ||
</div> | </div> |
Revision as of 03:40, 6 October 2021
![](https://static.igem.org/mediawiki/2021/2/27/T--Shanghai_United--homepage01.png)
![](https://static.igem.org/mediawiki/2021/a/a5/T--Shanghai_United--Description00.png)
Notebook
Jul. 28, 2021
- Learnt and clarified research goals and methods as a team. Learnt safety precautions necessary for entering the laboratory.
- Learnt and practiced the use of micropipettes.
- Performed PCR of amilGFP gene; performed gel electrophoresis of said PCR products and tested for results.
- Researched and learnt the reaction mechanisms of PCR and the preparation protocol of LB cultivation medium.
- Used LB cultivation medium with KANA antibiotics to cultivate three E. coli cell lines that are KANA resistant and with ArsA, ArsD, and ArsR genes inserted, respectively.
Jul. 29, 2021
- Performed plasmid extraction of three E. coli cell lines, cultivated since Jul. 28. Said bacteria have KANA resistance and ArsA, ArsD, or ArsR depending on the cell line. Extracted plasmids are tested for and yielded desirable concentrations.
- Cleaned up amilGFP PCR products, obtained in Jul. 28. Cleaned up PCR products are tested for and yielded desirable concentrations.
- Using restriction enzyme BsaI, digested ArsA, ArsD, and ArsR plasmids. Reaction mix is left overnight to react.
- Using restriction enzymes BsaI and DpnI, digested cleaned up amilGFP PCR products. Reaction mix is left overnight to react.
Jul. 30, 2021
- Harvested restriction enzyme digestion products of ArsA, ArsD, and ArsR plasmids as well as amilGFP PCR products; digestion was conducted in Jul. 29.
- Performed gel electrophoresis on restriction enzyme digestion products of ArsA, ArsD, ArsR, and amilGFP genes.
- Performed agarose gel DNA extraction of gel containing restriction enzyme digestion products. Extracted ArsA, ArsD, ArsR, and amilGFP genes are tested for and yielded suboptimal concentrations.
- Performed T4 DNA ligase ligation of Ars genes and amilGFP genes.
- Using ligation products of Ars genes and amilGFP genes, transformed DH5-α E. coli by heat shock method.
- Prepared LB cultivation medium agar plates with 0.1% KANA antibiotics.
- Spread Ars/amilGFP E. coli onto KANA antibiotics agar plates.
Jul. 31, 2021
Gel Electrophoresis Results of amilGFP PCR
Gel Electrophoresis Results of Colony PCR
- 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.
- Performed colony PCR and gel electrophoresis to test for Ars/amilGFP genes in observed E. coli colonies. Only one colony resulted to be ArsD/amilGFP positive, indicating failure of experimental procedure.
- Performed amilGFP restriction enzyme digestion on cleaned up amilGFP PCR products, obtained in Jul. 29, to prepare for first retry of experimental procedure.
- Performed PCR of amilGFP gene to prepare for second retry of experimental procedure. Performed electrophoresis to test for amilGFP PCR products.
- Performed gel electrophoresis to test for amilGFP restriction enzyme digestion products. Performed agarose gel DNA extraction for digested amilGFP genes.
- Cleaned up amilGFP PCR products.
- Performed T4 DNA ligase ligation of digested amilGFP genes and digested Ars genes, obtained in Jul. 30.
- Cultivated three cell lines of E. coli with KANA resistance and ArsA, ArsD, or ArsR genes inserted, depending on the cell line, to prepare for second retry of experimental procedure.
- Using ligation products of Ars genes and amilGFP genes, transformed DH5-α E. coli by heat shock method.
- Spread Ars/amilGFP E. coli onto KANA antibiotics agar plates, prepared in Jul. 30.
Aug. 1, 2021
- Examined growths of Ars/amilGFP E. coli, cultivated since Jul. 31. Results were optimal: colonies were observed on all six plates.
- Performed colony PCR and gel electrophoresis to test for Ars/amilGFP genes in observed E. coli colonies. All selected colonies yielded positive results of Ars genes.
- Extracted ArsA, ArsD, and ArsR plasmids from Ars E. coli, cultivated Jul. 31. Because the first retry of experimental procedure yielded positive results, the second retry had been terminated.
- Cultivated Ars/amilGFP positive E. coli.
Aug. 2, 2021
- Preserved Ars/amilGFP positive E. coli for future studies.
- Conducted function test of Ars/amilGFP positive E. coli with different concentrations of NaH2PO4 solutions.
- Mailed Ars/amilGFP positive E. coli samples to sequencing companies to perform Sanger sequencing of Ars/amilGFP genes.
- Started construction of team Wikipedia reports.
Aug. 3, 2021
- Examined function test results, conducted in Aug. 2. No green fluorescence was found in all tubes.
- Conducted second function test of Ars/amilGFP positive E. coli with higher concentrations of NaH2PO4.
- Continued construction of team Wikipedia.
- Created product design information and user manual sheet for dry team for reference.
Aug. 4, 2021
- Examined second function test results, conducted in Aug. 3. No green fluorescence was found in all tubes, indicating the inability of Ars/amilGFP E. coli to detect phosphate.
- Analyzed results of Ars/amilGFP gene sequencing results, sequenced in Aug. 2. Constructed Ars/amilGFP genes are correct.
- Conducted third function test of Ars/amilGFP positive E. coli with different concentrations of C2H6AsNaO2.
CONTACT INFO
Please scan the following QR codes to find more about us on
Weibo、Bilibili、Wechat !
![](https://static.igem.org/mediawiki/2021/1/1d/T--Shanghai_United--homepage06.jpg)