Difference between revisions of "Team:Shanghai high school/Notebook"

(Prototype team page)
 
Line 1: Line 1:
−
{{IGEM_TopBar}}
 
−
{{Shanghai_high_school}}
 
 
<html>
 
<html>
  
−
<div class="column full_size">
+
<link rel="stylesheet" type="text/css"
 +
      href="https://2021.igem.org/wiki/index.php?title=Template:Shanghai_high_school/StyleCSS&action=raw&ctype=text/css"/>
  
−
<h1>Notebook</h1>
+
<div class="dr-oral-content">
−
<p> Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.</p>
+
    <!--navbar-->
 +
    <div class="navbar">
 +
        <a class="logo" href="https://2021.igem.org/Team:Shanghai_high_school">
 +
            <img src="https://static.igem.org/mediawiki/2021/7/7f/T--Shanghai_high_school--logo.png" alt="">
 +
        </a>
  
−
</div>
+
        <!--nav-->
−
<div class="clear"></div>
+
        <ul class="list">
 +
            <!--Team-->
 +
            <li class="item">
 +
                <a href="https://2021.igem.org/Team:Shanghai_high_school/Team">Team</a>
 +
                <div class="child-nav">
 +
                    <ul class="child-list">
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Team">Team Members</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Attributions">Attributions</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Collaborations">Collaboration</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Partnership">Partnership</a>
 +
                        </li>
 +
                    </ul>
 +
                </div>
 +
            </li>
  
 +
            <!--Modeling-->
 +
            <li class="item">
 +
                <a href="https://2021.igem.org/Team:Shanghai_high_school/Model">Modeling</a>
 +
            </li>
  
 +
            <!--Entrepreneurship-->
 +
            <li class="item">
 +
                <a href="https://2021.igem.org/Team:Shanghai_high_school/Entrepreneurship">Entrepreneurship</a>
 +
            </li>
  
−
<div class="column two_thirds_size">
+
            <!--Implementation-->
−
<h3>What should this page have?</h3>
+
            <li class="item">
−
<ul>
+
                <a href="https://2021.igem.org/Team:Shanghai_high_school/Implementation">Implementation</a>
−
<li>Chronological notes of what your team is doing.</li>
+
            </li>
−
<li> Brief descriptions of daily important events.</li>
+
−
<li>Pictures of your progress. </li>
+
−
<li>Mention who participated in what task.</li>
+
−
</ul>
+
  
−
</div>
+
            <!--Human Practice-->
 +
            <li class="item">
 +
                <a href="https://2021.igem.org/Team:Shanghai_high_school/Human_Practices">Human Practice</a>
 +
                <div class="child-nav">
 +
                    <ul class="child-list">
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Human_Practices">Integrated Human
 +
                                Practice</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Communication">Communication</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Fundraising">Fundraising</a>
 +
                        </li>
 +
                    </ul>
 +
                </div>
 +
            </li>
  
−
<div class="column third_size">
+
            <!--Parts-->
−
<div class="highlight decoration_A_full">
+
            <li class="item">
−
<h3>Inspiration</h3>
+
                <a href="https://2021.igem.org/Team:Shanghai_high_school/PartsCollection">Parts</a>
−
<p>You can see what others teams have done to organize their notes:</p>
+
                <div class="child-nav">
 +
                    <ul class="child-list">
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Parts_Collection">Parts
 +
                                Collection</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Engineering">Engineering</a>
 +
                        </li>
 +
                    </ul>
 +
                </div>
 +
            </li>
  
−
<ul>  
+
            <!--Project-->
−
<li><a href="https://2018.igem.org/Team:Munich/Notebook">2018 Munich</a></li>
+
            <li class="item active">
 +
                <a href="https://2021.igem.org/Team:Shanghai_high_school/Description">Project</a>
 +
                <div class="child-nav">
 +
                    <ul class="child-list">
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Description">Description</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Experiments">Experiments</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Results">Result</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/ProofOfConcept">Proof of Concept</a>
 +
                        </li>
 +
                        <li class="child-item active">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Notebook">Notebook</a>
 +
                        </li>
 +
                        <li class="child-item">
 +
                            <a href="https://2021.igem.org/Team:Shanghai_high_school/Safety">Safety</a>
 +
                        </li>
 +
                    </ul>
 +
                </div>
 +
            </li>
  
−
<li><a href="https://2019.igem.org/Team:Georgia_State/Notebook">2019 Georgia State</a></li>
+
            <!--Home-->
−
<li><a href="https://2019.igem.org/Team:Newcastle/Notebook">2019 Newcastle</a></li>
+
            <li class="item">
 +
                <a href="https://2021.igem.org/Team:Shanghai_high_school">Home</a>
 +
            </li>
 +
        </ul>
 +
    </div>
  
 +
    <!--图片-->
 +
    <div class="page-header">
 +
        <img src="https://static.igem.org/mediawiki/2021/f/fd/T--Shanghai_high_school--bg_1.png" alt="">
 +
        <h1>Notebook</h1>
 +
    </div>
  
−
<li><a href="https://2020.igem.org/Team:IISER-Pune-India/Notebook">2020 IISER Pune India</a></li>
+
    <!--内容-->
−
<li><a href="https://2020.igem.org/Team:Lund/Notebook">2020 Lund</a></li>
+
    <div class="content-small">
−
<li><a href="https://2020.igem.org/Team:NOVA_LxPortugal/Notebook">2020 NOVA LxPortugal</a></li>
+
        <section class="article p-t-54 p-b-54">
−
<li><a href="https://2020.igem.org/Team:RDFZ-China/NoteBook">2020 RDFZ China</a></li>
+
            <section>
−
</ul>
+
                <h1 class="title">Online: Online Training, Topic Discussion - June~July</h1>
−
</div>
+
            </section>
−
</div>
+
  
 +
            <section>
 +
                <h1 class="title">Offline:</h1>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 1 2021/08/05</h2>
 +
 +
                    <ul class="normal">
 +
                        <li><p>Ice breaking: self-introduction & games</p></li>
 +
                        <li><p>Elected 3 preliminary leaders for dry team and 2 for wet team</p></li>
 +
                        <li><p>Designed team name and first drafts of logo and uniform, decided the types of peripheral
 +
                            products</p></li>
 +
                        <li><p>Introduction of dry team coach and tasks of the day and after</p></li>
 +
                        <li><p>Everyone wrote their self-introductions</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12" src="https://static.igem.org/mediawiki/2021/8/81/T--Shanghai_high_school--img_notebook_1.jpg" alt="" style="width: 90%">
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 2 2021/08/06</h2>
 +
 +
                    <ul class="normal">
 +
                        <li><p>Laboratory safety training, instrument introduction</p></li>
 +
                        <li><p>Established experimental design framework and introduced basic experimental principles</p>
 +
                        </li>
 +
                        <li><p>Prepared LB solid / liquid culture medium and poured Amp / Kan antibiotics to the plates</p>
 +
                        </li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/c/cd/T--Shanghai_high_school--img_notebook_2.jpg" alt=""
 +
                        style="width: 90%">
 +
 +
                    <ul class="normal">
 +
                        <li><p>Prepared competent solutions for <i>Bacillus subtilis</i>: 200 mL (LB + 0.5 M Sorbitol), 200
 +
                            mL electroporation culture medium, and 10 mL RM</p></li>
 +
                        <li><p>Plasmid transformation: transferred pET28a(+) and PHT43-His into competent <i>E. coli</i> DH5
 +
                            α</p></li>
 +
                        <li><p>Tried to re-activate <i>Bacillus subtilis</i> strain WB800n and DH5α carrying pET28a plasmid
 +
                        </p></li>
 +
                    </ul>
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 3 2021/08/07</h2>
 +
 +
                    <ul class="normal">
 +
                        <li><p>Transformed <i>Bacillus subtilis</i> plasmid WB800n into competent cells of <i>E. coli</i>
 +
                        </p></li>
 +
                        <li><p>Extracted and purified pET28a plasmid, which was digested overnight with enzyme BamHI</p>
 +
                        </li>
 +
                        <li><p>Inoculated the re-activated <i>Bacillus subtilis</i> into the culture medium for competence
 +
                            preparation and cultured it overnight</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12" src="https://static.igem.org/mediawiki/2021/5/5c/T--Shanghai_high_school--img_notebook_3.jpg" alt="" style="width: 90%">
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 4 2021/08/08</h2>
 +
 +
                    <ul class="normal">
 +
                        <li><p>Successfully prepared <i>Bacillus subtilis</i> competent cells </p></li>
 +
                        <li><p>Transformed pET28a-VP7-LTB into <i>E. coli</i> competent cell line BL21 (DE3)</p></li>
 +
                        <li><p>Small scale of PHT43-His plasmid extraction</p></li>
 +
                        <li><p>Used PCR to amplify the DNA sequence of VP7 and VP7-LTB, confirmed with agarose gel
 +
                            electrophoresis, sent to a company to be sequenced</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/5/5b/T--Shanghai_high_school--img_notebook_4.jpg" alt=""
 +
                        style="width: 90%">
 +
 +
                    <ul class="normal">
 +
                        <li><p>First round PCR and second round PCR (did the second round twice)</p></li>
 +
                        <li><p>Recovered enzyme digestion linearization vectors by gel-cutting</p></li>
 +
                    </ul>
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 5 2021/08/09</h2>
 +
 +
                    <ul class="normal">
 +
                        <li><p>PHT43-His plasmid small extraction</p></li>
 +
                        <li><p>Electroporation: transferred PHT43-His plasmid to <i>Bacillus subtilis</i> to detect whether
 +
                            the competence preparation and electroporation system are feasible</p></li>
 +
                        <li><p>Prepared of <i>E. coli</i> and Bacillus subtilis induced expression culture medium (TM
 +
                            medium)</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/4/42/T--Shanghai_high_school--img_notebook_5.jpg" alt=""
 +
                        style="width: 90%">
 +
 +
                    <ul class="normal">
 +
                        <li><p>Recovered VP7 fragment and VP7-LTB by gel cutting</p></li>
 +
                    </ul>
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 6 2021/08/10</h2>
 +
 +
                    <p>One day off</p>
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 7 2021/08/11</h2>
 +
 +
                    <ul class="normal">
 +
                        <li><p>Optimized primers, completed PCR amplification of the target gene of each expression
 +
                            vector</p></li>
 +
                        <li><p>Transformed pET28a non-loaded plasmid into <i>E. coli</i> competent cells as a negative
 +
                            control for induced expression</p></li>
 +
                        <li><p>IPTG induced expression in <i>E. coli</i>, examined ITPG concentration</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/4/42/T--Shanghai_high_school--img_notebook_6.jpg" alt=""
 +
                        style="width: 90%">
 +
 +
                    <ul class="normal">
 +
                        <li><p>Explored the use of the ultrasonic crusher</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/b/b1/T--Shanghai_high_school--img_notebook_7.jpg" alt=""
 +
                        style="width: 90%">
 +
 +
                    <ul class="normal">
 +
                        <li><p>Plasmid digestion, homologous recombination, vector construction</p></li>
 +
                        <li><p>Explained the experimental principles and precautions of SDS-PAGE and Western-blot</p></li>
 +
                        <li><p>Expanded the culture of positive monoclonal antibody of <i>Bacillus subtilis</i> and tried
 +
                            colony PCR to further eliminate false positive</p></li>
 +
                    </ul>
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 8 2021/08/12</h2>
 +
 +
                    <ul class="normal">
 +
                        <li><p>Inducible expression of VP7-LTB growing in BL21 using IPTG, establish bacterial growth curve
 +
                            by measuring OD<sub>600</sub>, and collect samples with different IPTG concentration at
 +
                            different time (0, 2, 4, 5, and 6 hrs)</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/5/5f/T--Shanghai_high_school--img_notebook_8.jpg" alt=""
 +
                        style="width: 50%">
 +
 +
                    <ul class="normal">
 +
                        <li><p>Optimize the electroporation system of <i>B. Subtilis</i></p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/b/b3/T--Shanghai_high_school--img_notebook_9.jpg" alt=""
 +
                        style="width: 70%">
 +
 +
                    <ul class="normal">
 +
                        <li><p>SDS-PAGE gel preparation </p></li>
 +
                        <li><p>construct plasmids with target genes: pET28a-VP7, PHT43-His-VP7, PHT43-His-VP7-LTB, using
 +
                            homologous recombination</p></li>
 +
                    </ul>
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 9 2021/08/13</h2>
 +
 +
                    <ul class="normal">
 +
                        <li><p>Extraction of protein of VP7-LTB</p></li>
 +
                        <li><p>SDS-PAGE preparation</p></li>
 +
                        <li><p>Establish bacterial growth curve by measuring OD<sub>600</sub></p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/b/bc/T--Shanghai_high_school--img_notebook_10.jpg" alt=""
 +
                        style="width: 70%">
 +
 +
                    <ul class="normal">
 +
                        <li><p><i>B. Subtilis</i> inducible expression preliminary experiment</p></li>
 +
                        <li><p>Expansion culture of pET28a-VP7，PHT43-his-VP7，PHT43-his-VP7-LTB, and DNA purification for
 +
                            transformation</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/e/ec/T--Shanghai_high_school--img_notebook_11.jpg" alt=""
 +
                        style="width: 70%">
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 10 2021/08/14</h2>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/e/e6/T--Shanghai_high_school--img_notebook_12.jpg" alt="" style="width: 70%">
 +
 +
                    <ul class="normal">
 +
                        <li><p>SDS-PAGE: VP7-LTB 1nM IPTG</p></li>
 +
                        <li><p>Establishment of all carriers </p></li>
 +
                        <li><p>Inducible expression of VP7 in BL21 using IPTG</p></li>
 +
                        <li><p><i>B. Subtilis</i> electroporation</p></li>
 +
                        <li><p>Personal and group photo</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/1/10/T--Shanghai_high_school--img_notebook_13.jpg" alt="" style="width: 70%">
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 11 2021/08/15</h2>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/9/9e/T--Shanghai_high_school--img_notebook_14.jpg" alt="" style="width: 70%">
 +
 +
                    <ul class="normal">
 +
                        <li><p>SDS-PAGE: VP7-LTB 0.5 nM, 2 nM IPTG, VP7 1nM IPTG</p></li>
 +
                        <li><p>Observe the positive single colony of WB800n</p></li>
 +
                        <li><p>Summary of the project</p></li>
 +
                        <li><p>Presentation preparation: power point</p></li>
 +
                    </ul>
 +
 +
                </section>
 +
 +
                <section>
 +
                    <h2 class="title2">Day 12 2021/08/16</h2>
 +
 +
                    <ul class="normal">
 +
                        <li><p>Periodic report</p></li>
 +
                    </ul>
 +
 +
                    <img class="m-t-12 m-b-12" src="https://static.igem.org/mediawiki/2021/0/09/T--Shanghai_high_school--img_notebook_15.jpg" alt="" style="width: 90%">
 +
                </section>
 +
            </section>
 +
        </section>
 +
    </div>
 +
 +
    <!--Footer-->
 +
    <div class="footer">
 +
        <div class="content">
 +
            <hr>
 +
            <span>Contact Email: 1649407718@qq.com; 2796437537@qq.com</span>
 +
            <span>Wechat Search ‘Dr Oral’ and follow us!</span>
 +
        </div>
 +
    </div>
 +
</div>
 
</html>
 
</html>

Revision as of 16:20, 23 September 2021

Notebook

Online: Online Training, Topic Discussion - June~July

Offline:

Day 1 2021/08/05

  • Ice breaking: self-introduction & games

  • Elected 3 preliminary leaders for dry team and 2 for wet team

  • Designed team name and first drafts of logo and uniform, decided the types of peripheral products

  • Introduction of dry team coach and tasks of the day and after

  • Everyone wrote their self-introductions

Day 2 2021/08/06

  • Laboratory safety training, instrument introduction

  • Established experimental design framework and introduced basic experimental principles

  • Prepared LB solid / liquid culture medium and poured Amp / Kan antibiotics to the plates

  • Prepared competent solutions for Bacillus subtilis: 200 mL (LB + 0.5 M Sorbitol), 200 mL electroporation culture medium, and 10 mL RM

  • Plasmid transformation: transferred pET28a(+) and PHT43-His into competent E. coli DH5 α

  • Tried to re-activate Bacillus subtilis strain WB800n and DH5α carrying pET28a plasmid

Day 3 2021/08/07

  • Transformed Bacillus subtilis plasmid WB800n into competent cells of E. coli

  • Extracted and purified pET28a plasmid, which was digested overnight with enzyme BamHI

  • Inoculated the re-activated Bacillus subtilis into the culture medium for competence preparation and cultured it overnight

Day 4 2021/08/08

  • Successfully prepared Bacillus subtilis competent cells

  • Transformed pET28a-VP7-LTB into E. coli competent cell line BL21 (DE3)

  • Small scale of PHT43-His plasmid extraction

  • Used PCR to amplify the DNA sequence of VP7 and VP7-LTB, confirmed with agarose gel electrophoresis, sent to a company to be sequenced

  • First round PCR and second round PCR (did the second round twice)

  • Recovered enzyme digestion linearization vectors by gel-cutting

Day 5 2021/08/09

  • PHT43-His plasmid small extraction

  • Electroporation: transferred PHT43-His plasmid to Bacillus subtilis to detect whether the competence preparation and electroporation system are feasible

  • Prepared of E. coli and Bacillus subtilis induced expression culture medium (TM medium)

  • Recovered VP7 fragment and VP7-LTB by gel cutting

Day 6 2021/08/10

One day off

Day 7 2021/08/11

  • Optimized primers, completed PCR amplification of the target gene of each expression vector

  • Transformed pET28a non-loaded plasmid into E. coli competent cells as a negative control for induced expression

  • IPTG induced expression in E. coli, examined ITPG concentration

  • Explored the use of the ultrasonic crusher

  • Plasmid digestion, homologous recombination, vector construction

  • Explained the experimental principles and precautions of SDS-PAGE and Western-blot

  • Expanded the culture of positive monoclonal antibody of Bacillus subtilis and tried colony PCR to further eliminate false positive

Day 8 2021/08/12

  • Inducible expression of VP7-LTB growing in BL21 using IPTG, establish bacterial growth curve by measuring OD600, and collect samples with different IPTG concentration at different time (0, 2, 4, 5, and 6 hrs)

  • Optimize the electroporation system of B. Subtilis

  • SDS-PAGE gel preparation

  • construct plasmids with target genes: pET28a-VP7, PHT43-His-VP7, PHT43-His-VP7-LTB, using homologous recombination

Day 9 2021/08/13

  • Extraction of protein of VP7-LTB

  • SDS-PAGE preparation

  • Establish bacterial growth curve by measuring OD600

  • B. Subtilis inducible expression preliminary experiment

  • Expansion culture of pET28a-VP7，PHT43-his-VP7，PHT43-his-VP7-LTB, and DNA purification for transformation

Day 10 2021/08/14

  • SDS-PAGE: VP7-LTB 1nM IPTG

  • Establishment of all carriers

  • Inducible expression of VP7 in BL21 using IPTG

  • B. Subtilis electroporation

  • Personal and group photo

Day 11 2021/08/15

  • SDS-PAGE: VP7-LTB 0.5 nM, 2 nM IPTG, VP7 1nM IPTG

  • Observe the positive single colony of WB800n

  • Summary of the project

  • Presentation preparation: power point

Day 12 2021/08/16

  • Periodic report

Contact Email: 1649407718@qq.com; 2796437537@qq.com Wechat Search ‘Dr Oral’ and follow us!