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| | {{:Team:TUDelft/Banner}} | | {{:Team:TUDelft/Banner}} |
| | <html> | | <html> |
| − | | + | <body> |
| − | <head>
| + | |
| − | <meta name="viewport" content="width=device-width, initial-scale=1">
| + | |
| − | <style>
| + | |
| − | | + | |
| − | #tabletu {
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| − | background-color: transparent;
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| − | border-collapse: collapse;
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| − | width:100%;
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| − | }
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| − | | + | |
| − | #tabletu td, th {
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| − | border: 1px solid #000000;
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| − | padding: 8px;
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| − | }
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| − | | + | |
| − | #tabletu th {
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| − | padding: 8px;
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| − | text-align: left;
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| − | border: 1px solid #000000;
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| − | background-color: rgba(0,110,167,1);
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| − | color: white;
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| − | }
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| + | |
| − | #tabletu td {
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| − | height:48px;
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| − | }
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| − | .modalTD {padding: 0!important;
| + | |
| − | background-color: #00a6d6;
| + | |
| − | }
| + | |
| − |
| + | |
| − | /* The Modal (background) */
| + | |
| − | .modal {
| + | |
| − | display: none; /* Hidden by default */
| + | |
| − | position: fixed; /* Stay in place */
| + | |
| − | z-index: 1; /* Sit on top */
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| + | |
| − | overflow: auto; /* Enable scroll if needed */
| + | |
| − | background-color: rgb(0,0,0); /* Fallback color */
| + | |
| − | background-color: rgba(0,0,0,0.4); /* Black w/ opacity */
| + | |
| − | }
| + | |
| − | | + | |
| − | /* Modal Content */
| + | |
| − | .modal-content {
| + | |
| − | position: relative;
| + | |
| − | background-color: #fefefe;
| + | |
| − | margin: auto;
| + | |
| − | padding: 0px;
| + | |
| − | border: 1px solid #888;
| + | |
| − | width: 80%;
| + | |
| − | box-shadow: 0 4px 8px 0 rgba(0,0,0,0.2),0 6px 20px 0 rgba(0,0,0,0.19);
| + | |
| − | -webkit-animation-name: animatetop;
| + | |
| − | -webkit-animation-duration: 0.4s;
| + | |
| − | animation-name: animatetop;
| + | |
| − | animation-duration: 0.4s
| + | |
| − | }
| + | |
| − | | + | |
| − | /* Add Animation */
| + | |
| − | @-webkit-keyframes animatetop {
| + | |
| − | from {top:-300px; opacity:0}
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| − | to {top:0; opacity:1}
| + | |
| − | }
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| − | | + | |
| − | @keyframes animatetop {
| + | |
| − | from {top:-300px; opacity:0}
| + | |
| − | to {top:0; opacity:1}
| + | |
| − | }
| + | |
| − | | + | |
| − | /* The Close Button */
| + | |
| − | .close {
| + | |
| − | color: black;
| + | |
| − | float: right;
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| − | font-size: 28px;
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| − | font-weight: bold;
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| − | }
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| − | .close:hover,
| + | |
| − | .close:focus {
| + | |
| − | color: white;
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| − | text-decoration: none;
| + | |
| − | cursor: pointer;
| + | |
| − | }
| + | |
| − | | + | |
| − | .notebook {
| + | |
| − | border-collapse: collapse;
| + | |
| − | width:70%;}
| + | |
| − | | + | |
| − | .notebook td.modalTD:hover .block:hover {background-color: yellow;
| + | |
| − | color: black;}
| + | |
| − | .notebook td, .notebook th {
| + | |
| − | border: 1px solid #ddd;
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| − | padding: 0px;
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| − | }
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| − | .block {
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| − | display: block;
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| − | width: 100%;
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| − | height:100%;
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| − | border: none;
| + | |
| − | background-color: #00a6d6;
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| − | padding: 14px 28px;
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| − | font-size: 16px;
| + | |
| − | cursor: pointer;
| + | |
| − | text-align: center;
| + | |
| − | box-sizing: border-box;
| + | |
| − | }
| + | |
| − | .month {
| + | |
| − | text-align: center;
| + | |
| − | font-weight: bold;
| + | |
| − | }
| + | |
| − | #category {
| + | |
| − | padding:8px;
| + | |
| − | background-color: transparent;
| + | |
| − | }
| + | |
| − | .modal-header {
| + | |
| − | padding: 30px 30px;
| + | |
| − | background-color: yellow ;
| + | |
| − |
| + | |
| − | }
| + | |
| − | .modal-body {padding: 20px 16px;
| + | |
| − | overflow-y: auto;
| + | |
| − | height:450px;
| + | |
| − | }
| + | |
| − | | + | |
| − | .modal-footer {
| + | |
| − | padding: 30px 16px;
| + | |
| − | background-color: yellow;
| + | |
| − | }
| + | |
| − | .modal-header h2 {
| + | |
| − | color:black;
| + | |
| − | }
| + | |
| − |
| + | |
| − | .modal-content table {
| + | |
| − | border-collapse: collapse;
| + | |
| − | width:100%;
| + | |
| − | }
| + | |
| − |
| + | |
| − | .modal-content tbody tr td:first-child {
| + | |
| − |
| + | |
| − | width: 17%;
| + | |
| − | }
| + | |
| − | .modal-content a:link {
| + | |
| − | color:black;
| + | |
| − | text-decoration: none;
| + | |
| − | }
| + | |
| − | .modal-content a:hover {
| + | |
| − | color: #00A6D6;
| + | |
| − | }
| + | |
| − | </style>
| + | |
| − | </head>
| + | |
| − | <body>
| + | |
| | <div class="Banner container-fluid text-center mb-0 align-items-center "> | | <div class="Banner container-fluid text-center mb-0 align-items-center "> |
| | | | |
| Line 166: |
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| | <br> | | <br> |
| | <br> | | <br> |
| − | <img src = "https://static.igem.org/mediawiki/2019/e/e4/T--TUDelft--Notebook_logo.png" alt="Design" style="width:60%";> | + | <img src = "https://static.igem.org/mediawiki/2019/e/e4/T--TUDelft--Notebook_logo.png" alt="Notebook" style="width:60%";> |
| | | | |
| | </div> | | </div> |
| | </div> | | </div> |
| | </div> | | </div> |
| − | <div class="centerjustify">
| + | |
| − | <h2>Orthogonal Replication</h2>
| + | |
| − | <br>
| + | |
| − | <table id="tabletu" class="notebook">
| + | |
| − | <tr id="category">
| + | |
| − | <th style="background-color:transparent; color:black;"></th>
| + | |
| − | <th style="background-color:transparent; color:black;" colspan=2> Cloning</th>
| + | |
| − | <th style="background-color:transparent; color:black;" colspan=2>Testing of Plasmid</th>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td class="month">April</td>
| + | |
| − | <td rowspan=4 class="modalTD">
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal1">Gibson Assembly of Gene of Interest Plasmid</button>
| + | |
| − | | + | |
| − | <!-- The Modal -->
| + | |
| − | <div id="myModal1" class="modal">
| + | |
| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
| + | |
| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>Gibson Assembly of Gene of Interest Plasmid</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Goal:</td>
| + | |
| − | <td>Construction of the Gene of Interest Plasmid</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Protocols Used:</td>
| + | |
| − | <td> <a href="https://2019.igem.org/Team:TUDelft/Experiments">Nanodrop DNA quantification</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Plasmid Isolation</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Phusion PCR</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Restriction Digestion</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E.coli</i> Heat Shock Transformation</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">KOD Extreme PCR</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Colony Polymerase PCR (GoTaq)</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E. coli</i> Glycerol Stock</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">Gibson Assembly</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">DNA Gel Purification</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">DNA Gel Electrophoresis</a>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Plan:</td>
| + | |
| − | <td>The plasmid we had contained OriL and OriR flanking GFP. We wanted to clone a kanamycin gene inside this to use as a selection marker.</td></tr>
| + | |
| − | <tr>
| + | |
| − | <td>Results:
| + | |
| − | </td>
| + | |
| − | <td>There were no plates with colonies, therefore Gibson Assembly has not work.</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | <div class="modal-footer">
| + | |
| − |
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </td>
| + | |
| − | <td rowspan=4 class="modalTD" >
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal2">Gibson Assembly of Support Plasmid</button>
| + | |
| − | | + | |
| − | <!-- The Modal -->
| + | |
| − | <div id="myModal2" class="modal">
| + | |
| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
| + | |
| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>Gibson Assembly of Support Plasmid</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Goal:</td>
| + | |
| − | <td>Construction of the Support Plasmid</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Protocols Used:</td>
| + | |
| − | <td> <a href="https://2019.igem.org/Team:TUDelft/Experiments">Nanodrop DNA quantification</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Plasmid Isolation</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Phusion PCR</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Restriction Digestion</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E.coli</i> Heat Shock Transformation</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">KOD Extreme PCR</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Colony Polymerase PCR (GoTaq)</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E. coli</i> Glycerol Stock</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">Gibson Assembly</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">DNA Gel Purification</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">DNA Gel Electrophoresis</a>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Plan:</td>
| + | |
| − | <td>We had two different plasmids. One plasmid with p5 and p6 on it and one plasmid with phi 29 DNAP and phi 29 TP. We wanted to put this on one plasmid through Gibson Assembly</td></tr>
| + | |
| − | <tr>
| + | |
| − | <td>Results:
| + | |
| − | </td>
| + | |
| − | <td>There were no plates with colonies, therefore Gibson Assembly has not work.</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | <div class="modal-footer">
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </td>
| + | |
| − | <td colspan=2></td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td class="month">May</td>
| + | |
| − | | + | |
| − | | + | |
| − | <td colspan=2></td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td class="month">June</td>
| + | |
| − | <td colspan=2></td>
| + | |
| − | | + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td class="month">July</td>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <td colspan=2 class="modalTD">
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal3">Brute Force Experiments 1</button>
| + | |
| − | | + | |
| − | <!-- The Modal -->
| + | |
| − | <div id="myModal3" class="modal">
| + | |
| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
| + | |
| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>Brute Force Experiments 1</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Goal:</td>
| + | |
| − | <td>Putting all separate plasmids in and see what survives</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Protocols Used:</td>
| + | |
| − | <td> <a href="https://2019.igem.org/Team:TUDelft/Experiments">Transformation</a>,
| + | |
| − | | + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">IPTG stock</a>,
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Plan:</td>
| + | |
| − | <td>As construction of the plasmids did not work, we just wanted to put all the separate constructs in, to see whether something would grow or not. We induced it with no IPTG, 10 µM IPTG and 50 µM IPTG. We put in the following constructs:
| + | |
| − | <ul>
| + | |
| − | <li>p02+p04+oriL-GFP-oriR</li>
| + | |
| − | <li>p02+oriL-GFP-oriR</li>
| + | |
| − | <li>oriL-GFP-oriR</li>
| + | |
| − | <li>p01+p02+p04</li>
| + | |
| − | </ul></td></tr>
| + | |
| − | <tr>
| + | |
| − | <td>Results:
| + | |
| − | </td>
| + | |
| − | <td>Some plates grew, some not. No conclusive result.</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | <div class="modal-footer">
| + | |
| − |
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td class="month">August</td>
| + | |
| − | <td rowspan=2 colspan=2 class="modalTD">
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal4">MoClo Assembly</button>
| + | |
| − | | + | |
| − | <!-- The Modal -->
| + | |
| − | <div id="myModal4" class="modal">
| + | |
| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
| + | |
| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>MoClo Assembly</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Goal:</td>
| + | |
| − | <td>Assembly all the different parts with MoClo</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Protocols Used:</td>
| + | |
| − | <td> <a href="https://2019.igem.org/Team:TUDelft/Experiments">Nanodrop DNA quantification</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Plasmid Isolation</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">MoClo</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E.coli</i> Heat Shock Transformation</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">KOD Extreme PCR</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Colony Polymerase PCR (GoTaq)</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E. coli</i> Glycerol Stock</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">DNA Gel Electrophoresis</a>
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">Sequencing</a>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Plan:</td>
| + | |
| − | <td>For Orthogonal Replication, first, all the level 0's were constructed and then assembled in level 1 to make the following constructs:
| + | |
| − | <ul>
| + | |
| − | <li>0.1 T7 p - U RBS - DNAP - WT</li>
| + | |
| − | <li>0.5 T7 p - U RBS - DNAP - WT</li> | + | |
| − | <li>WT T7 p - U RBS - DNAP - WT</li>
| + | |
| − | <li>0.1 T7 p - U RBS - TP - WT</li>
| + | |
| − | <li>0.5 T7 p - U RBS - TP - WT</li>
| + | |
| − | <li>WT T7 p - U RBS - TP - WT</li>
| + | |
| − | <li>0.1 T7 p - U RBS - p5 - WT</li>
| + | |
| − | <li>0.5 T7 p - U RBS - p5 - WT</li>
| + | |
| − | <li>WT T7 p - U RBS - p5 - WT</li>
| + | |
| − | <li>0.1 T7 p - U RBS - p6 - WT</li>
| + | |
| − | <li>0.5 T7 p - U RBS - p6 - WT</li>
| + | |
| − | <li>WT T7 p - U RBS - p6 - WT</li>
| + | |
| − | <li>0.1 T7 p - U RBS - GFP - WT</li>
| + | |
| − | <li>0.5 T7 p - U RBS - GFP - WT</li>
| + | |
| − | <li>WT T7 p - U RBS - GFP - WT</li>
| + | |
| − | </ul>
| + | |
| − |
| + | |
| − | </td></tr>
| + | |
| − | <tr>
| + | |
| − | <td>Results:
| + | |
| − | </td>
| + | |
| − | <td>All composite parts are confirmed by sequencing.</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | <div class="modal-footer">
| + | |
| − |
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </td>
| + | |
| − | <td colspan=2 class="modalTD">
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal5">Toxicity Experiments 1</button>
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| − | <div id="myModal5" class="modal">
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| − | <div class="modal-content">
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| − | <div class="modal-header">
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| − | <span class="close">×</span>
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| − | <h2>Toxicity Experiments 1</h2>
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| − | </div>
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| − | <div class="modal-body">
| + | |
| − | <table>
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| − | <tr>
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| − | <td>Goal:</td>
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| − | <td>Determinig what parts are toxic to the cell</td>
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| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Protocols Used:</td>
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| − | <td>
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| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E.coli</i> Heat Shock Transformation</a>,
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| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">OD600 Measurement</a>,
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| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">IPTG stock</a>,
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| − | </td>
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| − | </tr>
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| − | <tr>
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| − | <td>Plan:</td>
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| − | <td>
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| − | <ul>
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| − | <li>p01 (control)</li>
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| − | <li>p02</li>
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| − | <li>p03</li>
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| − | <li>p04</li>
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| − |
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| − | </ul>
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| − | </td></tr>
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| − | <tr>
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| − | <td>Results:
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| − | </td>
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| − | <td>They all died.</td>
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| − | </tr>
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| − | </table>
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| − | </div>
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| − | <div class="modal-footer">
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| − | <td rowspan=2 class="month">September</td>
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| − | <td rowspan=2 class="modalTD">
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| − | <!-- Trigger/Open The Modal -->
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| − | <button class="block modal-button" href="#myModal6">Toxicity Experiments 2</button>
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| − | <!-- The Modal -->
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| − | <div id="myModal6" class="modal">
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| − | <div class="modal-content">
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| − | <div class="modal-header">
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| − | <span class="close">×</span>
| + | |
| − | <h2>Toxicity Experiments 2</h2>
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| − | </div>
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| − | <div class="modal-body">
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Goal:</td>
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| − | <td>Determinig what parts are toxic to the cell</td>
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| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Protocols Used:</td>
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| − | <td>
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| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E.coli</i> Heat Shock Transformation</a>,
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| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">OD600 Measurement</a>,
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| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">IPTG stock</a>,
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| − | </td>
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| − | </tr>
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| − | <tr>
| + | |
| − | <td>Plan:</td>
| + | |
| − | <td>To determine which plasmids were toxic, we put transformed the plasmids separately in BL21 pLysS and set an overnight measurement to measure OD600 to see if the cells were alive.
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| − | <ul>
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| − | <li>p01 (control)</li>
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| − | <li>p02</li>
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| − | <li>p03</li>
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| − | <li>p04</li>
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| − |
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| − | </ul>
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| − | </td></tr>
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| − | <tr>
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| − | <td>Results:
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| − | </td>
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| − | <td>Add link</td>
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| − | </tr>
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| − | </table>
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| − | </div>
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| − | <div class="modal-footer">
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| − | </div>
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| − | </td>
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| − | <button class="block modal-button" href="#myModal7">Characterization of Level 1's</button>
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| − | <div class="modal-content">
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| − | <div class="modal-header">
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| − | <span class="close">×</span>
| + | |
| − | <h2>Characterization of Level 1's</h2>
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| − | </div>
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| − | <div class="modal-body">
| + | |
| − | <table>
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| − | <tr>
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| − | <td>Goal:</td>
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| − | <td>Add characterization for the parts page and see if the different promoters strength are visible</td>
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| − | </tr>
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| − | <tr>
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| − | <td>Protocols Used:</td>
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| − | <td>
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| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">PURE System</a>,
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| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">Protein Gel</a>,
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| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">Mass Spectrometry</a>,
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| − | </td>
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| − | </tr>
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| − | <tr>
| + | |
| − | <td>Plan:</td>
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| − | <td>The following constructs were added to the PURE system and then run on a protein gel:
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| − | <ul>
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| − | <li>0.1 T7 p - U RBS - DNAP - WT</li>
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| − | <li>0.5 T7 p - U RBS - DNAP - WT</li>
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| − | <li>WT T7 p - U RBS - DNAP - WT</li>
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| − | <li>0.1 T7 p - U RBS - TP - WT</li>
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| − | <li>0.5 T7 p - U RBS - TP - WT</li>
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| − | <li>WT T7 p - U RBS - TP - WT</li>
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| − | <li>0.1 T7 p - U RBS - p5 - WT</li>
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| − | <li>0.5 T7 p - U RBS - p5 - WT</li>
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| − | <li>WT T7 p - U RBS - p5 - WT</li>
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| − | <li>0.1 T7 p - U RBS - p6 - WT</li>
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| − | <li>0.5 T7 p - U RBS - p6 - WT</li>
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| − | <li>WT T7 p - U RBS - p6 - WT</li></ul>
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| − |
| + | |
| − | After this, a remainder of the sample was analyzed with Mass Spectrometry.
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| − | </td></tr>
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| − | <tr>
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| − | <td>Results:
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| − | </td>
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| − | <td>Add link</td>
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| − | </tr>
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| − | </table>
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| − | </div>
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| − | <div class="modal-footer">
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| − |
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| − | </div>
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| − | </div>
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| − | </div>
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| − | </td>
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| − | </tr>
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| − | <tr>
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| − | | + | |
| − | <td colspan=2></td>
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| − | | + | |
| − | </tr>
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| − | | + | |
| − | <tr>
| + | |
| − | <td class="month">October</td>
| + | |
| − | <td> </td>
| + | |
| − | <td> </td>
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| − | <td rowspan =2 class="modalTD">
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| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal8">Brute Force Experiments 2</button>
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| − | <!-- The Modal -->
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| − | <div id="myModal8" class="modal">
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| − | <!-- Modal content -->
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| − | <div class="modal-content">
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| − | <div class="modal-header">
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| − | <span class="close">×</span>
| + | |
| − | <h2>Brute Force Experiments 2</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Goal:</td>
| + | |
| − | <td>Putting all the needed components for orthogonal replication and see which cells replicate</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Protocols Used:</td>
| + | |
| − | <td> <a href="https://2019.igem.org/Team:TUDelft/Experiments">Transformation</a>,
| + | |
| − | | + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">IPTG stock</a>,
| + | |
| − | </td>
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| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Plan:</td>
| + | |
| − | <td>To test whether orthogonal replication works in vivo, we transformed all the plasmid with the components needed in pLysS together with a final construct, which is either OriL-Kan-OriR or OriL-GFP-Kan-OriR. This was done with different IPTG concentrations.
| + | |
| − |
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| − | <ul>
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| − | <li>oriL-Kan-OriR with
| + | |
| − | <ul>
| + | |
| − | <li>WT T7p TP, DNAP, p5, p6</li>
| + | |
| − | <li>0.5 T7p TP, DNAP, p5, p6</li>
| + | |
| − | <li>0.1 T7p TP, DNAP, p5, p6</li>
| + | |
| − | </ul>
| + | |
| − | </li>
| + | |
| − | <li>oriL-GFP-Kan-OriR
| + | |
| − | <ul>
| + | |
| − | <li>WT T7p TP, DNAP, p5, p6</li>
| + | |
| − | <li>0.5 T7p TP, DNAP, p5, p6</li>
| + | |
| − | <li>0.1 T7p TP, DNAP, p5, p6</li>
| + | |
| − | </ul></li>
| + | |
| − |
| + | |
| − | </ul></td></tr>
| + | |
| − | <tr>
| + | |
| − | <td>Results:
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| − | </td>
| + | |
| − | <td>To be done</td>
| + | |
| − | </tr>
| + | |
| − | </table>
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| − | </div>
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| − | <div class="modal-footer">
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| − |
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| − | </div>
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| − | </div>
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| − | </div>
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| − | </td>
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| − | <td rowspan =2 class="modalTD">
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal9">Orthogonal Replication</button>
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| − | | + | |
| − | <!-- The Modal -->
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| − | <div id="myModal9" class="modal">
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| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
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| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>Orthogonal Replication</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <p>Some text in the Modal Body</p>
| + | |
| − | <p>Some other text...</p>
| + | |
| − | </div>
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| − | <div class="modal-footer">
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| − | </div>
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| − | </tr>
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| − | </table>
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| − | <br><br>
| + | |
| − | <h2>Predictable and Transferable Expression</h2>
| + | |
| − | <br>
| + | |
| − | <table id="tabletu" class="notebook">
| + | |
| − | <tr id="category">
| + | |
| − | <th style="background-color:transparent; color:black;"></th>
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| − | <th style="background-color:transparent; color:black;">Cloning</th>
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| − | <th style="background-color:transparent; color:black;" colspan="3">Testing of Plasmid</th>
| + | |
| − | </tr>
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| − | <tr>
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| − | <td class="month">April</td>
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| − | <td></td>
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| − | <td colspan="3"></td>
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| − | </tr>
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| − | <tr>
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| − | <td class="month">May</td>
| + | |
| − | <td></td>
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| − | <td colspan="3"></td>
| + | |
| − | | + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td class="month">June</td>
| + | |
| − | <td class="modalTD" >
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal10">Uber Plasmid</button>
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| − | <!-- The Modal -->
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| − | <div id="myModal10" class="modal">
| + | |
| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
| + | |
| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>Uber Plasmid</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Goal:</td>
| + | |
| − | <td>Construction of the Uber Plasmid</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Protocols Used:</td>
| + | |
| − | <td> <a href="https://2019.igem.org/Team:TUDelft/Experiments">Nanodrop DNA quantification</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Plasmid Isolation</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Phusion PCR</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Restriction Digestion</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E.coli</i> Heat Shock Transformation</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Ligation</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">Colony Polymerase PCR (GoTaq)</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E. coli</i> Glycerol Stock</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">DNA Gel Purification</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">DNA Gel Electrophoresis</a>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Plan:</td>
| + | |
| − | <td>To digest the UBER plasmid and insert a casette with GFP to make the UBER plasmid fluorescence.</td></tr>
| + | |
| − | <tr>
| + | |
| − | <td>Results:
| + | |
| − | </td>
| + | |
| − | <td>Check PCR did not give the right size on the gel.</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | <div class="modal-footer">
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| − | </div>
| + | |
| − | </div>
| + | |
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| − | </div>
| + | |
| − | </td>
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| − | <td colspan="3"></td>
| + | |
| − | | + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td class="month">July</td>
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| − | | + | |
| − | <td></td>
| + | |
| − | <td colspan="3"></td>
| + | |
| − | | + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td class="month">August</td>
| + | |
| − | <td rowspan=3 class="modalTD">
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal11">MoClo Assembly</button>
| + | |
| − | | + | |
| − | <!-- The Modal -->
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| − | <div id="myModal11" class="modal">
| + | |
| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
| + | |
| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>MoClo Assembly</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Goal:</td>
| + | |
| − | <td>Assembly all the different parts with MoClo</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Protocols Used:</td>
| + | |
| − | <td> <a href="https://2019.igem.org/Team:TUDelft/Experiments">Nanodrop DNA quantification</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">Plasmid Isolation</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments">MoClo</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E.coli</i> Heat Shock Transformation</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">Colony Polymerase PCR (GoTaq)</a>, <a href="https://2019.igem.org/Team:TUDelft/Experiments"><i>E. coli</i> Glycerol Stock</a>,
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">DNA Gel Electrophoresis</a>
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">Sequencing</a>
| + | |
| − | <a href="https://2019.igem.org/Team:TUDelft/Experiments">Xgal Stock</a>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Plan:</td>
| + | |
| − | <td>For Predictable & Transferable Expression, first, all the level 0's were constructed and then assembled in level 1 to make the following constructs:
| + | |
| − | <ul>
| + | |
| − | <li>0.5 T7 p - SarJ + B0032 - TALEsp1 - WT T7 t</li>
| + | |
| − | <li>T7 p - SarJ + B0032 - TALEsp1 - WT T7 t</li>
| + | |
| − | <li>0.5 T7 p - RiboJ + B0032 - Juniper GFP - WT T7 t</li>
| + | |
| − | <li>T7_sp1 p - RiboJ + B0032 - Juniper GFP - WT T7 t</li>
| + | |
| − | <li>T7 p - RiboJ + B0032 - Juniper GFP - WT T7 t</li>
| + | |
| − | <li>0.5 T7_sp1 p - RiboJ + B0032 - Juniper GFP - WT T7 t</li>
| + | |
| − | <li>Pbhr - SarJ + B0032 - TALEsp1 - WT T7 t</li>
| + | |
| − | <li>Pbhr - RiboJ + B0032 - Juniper GFP - WT T7 t</li>
| + | |
| − | <li>0.5 T7 p - SarJ + B0032 - TALEsp1 - WT T7 t</li>
| + | |
| − | <li>T7 p - SarJ + B0032 - TALEsp1 - WT T7 t</li>
| + | |
| − | <li>Pbhr - Sarj + B0032 - TALEsp1 - WT T7 t</li>
| + | |
| − | <li>0.5 T7 p - RiboJ + B0032 - GFP - WT T7 t</li>
| + | |
| − | <li>0.1 T7 p - RiboJ + B0032 - GFP - WT T7 t</li>
| + | |
| − | <li>19bp T7 p - RiboJ + B0032 - GFP - WT T7 t</li>
| + | |
| − | <li>19bp 0.5 T7 p - RiboJ + B0032 - GFP - WT T7 t</li>
| + | |
| − | <li>19bp 0.1 T7 p - RiboJ + B0032 - GFP - WT T7 t</li>
| + | |
| − | <li>T7sp_sp1 p - RiboJ + B0032 - GFP - WT T7 t</li>
| + | |
| − | <li>Pbhr_sp1 - RiboJ + Universal RBS - GFP - WT T7 t</li>
| + | |
| − | <li>T7 p - SarJ + Universal RBS - GFP - WT T7 t</li>
| + | |
| − | | + | |
| − | </ul>
| + | |
| − | | + | |
| − | After assembling the level 1's, the following level M's were assembled:
| + | |
| − | <ul>
| + | |
| − | <li>T7p TALEsp1 - Dummy 2 - Dummy 3 - T7p TALEsp1</li>
| + | |
| − | <li>T7p TALEsp1 - Dummy 2 - Dummy 3 - T7p GFP</li>
| + | |
| − | <li>0.5 T7p TALEsp1 - Dummy 2 - Dummy 3 - 0.5 T7_sp1 p GFP</li>
| + | |
| − | <li>0.5 T7p TALEsp1 - Dummy 2 - Dummy 3 - 0.5 T7p GFP</li>
| + | |
| − | <li>0.5 T7p TALEsp1 - Dummy 2 - Dummy 3 - T7_sp1 p GFP</li>
| + | |
| − | <li>T7p TALEsp1 - Dummy 2 - Dummy 3 - 0.5 T7_sp1 p GFP</li>
| + | |
| − | <li>T7p TALEsp1 - T7p TALEsp1</li>
| + | |
| − | <li>T7p TALEsp1 - T7p GFP</li>
| + | |
| − | <li>0.5 T7p TALEsp1 - 0.5 T7_sp1 p GFP</li>
| + | |
| − | <li>T7p Universal RBS TALEsp1 - T7_sp1 p Universal RBS GFP</li>
| + | |
| − | <li>Pbhr Universal RBS TALEsp1 - Pbhr_sp1 Universal RBS GFP</li>
| + | |
| − | <li>Pbhr Universal RBS TALEsp1 - Pbhr_sp1 v2 Universal RBS GFP</li>
| + | |
| − | <li>Pbhr Universal RBS TALEsp1 - Pbhr Universal RBS GFP</li>
| + | |
| − | <li>Pbhr SarJ B00032 TALEsp1 - Pbhr Universal GFP</li>
| + | |
| − | </ul>
| + | |
| − | | + | |
| − | </td></tr>
| + | |
| − | <tr>
| + | |
| − | <td>Results:
| + | |
| − | </td>
| + | |
| − | <td>To be added</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | <div class="modal-footer">
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | </div>
| + | |
| − | <td colspan="3"></td>
| + | |
| − | | + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td class="month">September</td>
| + | |
| − |
| + | |
| − | <td rowspan=2 class="modalTD">
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal12">Copy Number Independence</button>
| + | |
| − | | + | |
| − | <!-- The Modal -->
| + | |
| − | <div id="myModal12" class="modal">
| + | |
| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
| + | |
| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>Copy Number Independence</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <p>Some text in the Modal Body</p>
| + | |
| − | <p>Some other text...</p>
| + | |
| − | </div>
| + | |
| − | <div class="modal-footer">
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </td>
| + | |
| − | <td rowspan=2 class="modalTD">
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal13">Transcriptional Variance Independence</button>
| + | |
| − | | + | |
| − | <!-- The Modal -->
| + | |
| − | <div id="myModal13" class="modal">
| + | |
| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
| + | |
| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>Transcriptional Variance Independence</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <p>Some text in the Modal Body</p>
| + | |
| − | <p>Some other text...</p>
| + | |
| − | </div>
| + | |
| − | <div class="modal-footer">
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </td>
| + | |
| − | <td rowspan=2 class="modalTD">
| + | |
| − | <!-- Trigger/Open The Modal -->
| + | |
| − | <button class="block modal-button" href="#myModal14">Translation Variance Independence</button>
| + | |
| − | | + | |
| − | <!-- The Modal -->
| + | |
| − | <div id="myModal14" class="modal">
| + | |
| − | | + | |
| − | <!-- Modal content -->
| + | |
| − | <div class="modal-content">
| + | |
| − | <div class="modal-header">
| + | |
| − | <span class="close">×</span>
| + | |
| − | <h2>Translation Variance Independence</h2>
| + | |
| − | </div>
| + | |
| − | <div class="modal-body">
| + | |
| − | <p>Some text in the Modal Body</p>
| + | |
| − | <p>Some other text...</p>
| + | |
| − | </div>
| + | |
| − | <div class="modal-footer">
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | </div>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − |
| + | |
| − | <tr>
| + | |
| − | <td class="month">October</td>
| + | |
| − |
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <script>
| + | |
| − | var btn = document.querySelectorAll("button.modal-button");
| + | |
| − | var modalTD = document.querySelectorAll(".modalTD") // get modalTD
| + | |
| − | | + | |
| − | | + | |
| − | var modals = document.querySelectorAll('.modal');
| + | |
| − | | + | |
| − | // Get the <span> element that closes the modal
| + | |
| − | var spans = document.getElementsByClassName("close");
| + | |
| − | | + | |
| − | // When the user clicks the button, open the modal
| + | |
| − | for (var i = 0; i < btn.length; i++) {
| + | |
| − | btn[i].style.height = modalTD[i].clientHeight + 'px' //set height of outer td to buttons
| + | |
| − | btn[i].onclick = function(e) {
| + | |
| − | e.preventDefault();
| + | |
| − | modal = document.querySelector(e.target.getAttribute("href"));
| + | |
| − | modal.style.display = "block";
| + | |
| − | }
| + | |
| − | }
| + | |
| − | | + | |
| − | // When the user clicks on <span> (x), close the modal
| + | |
| − | for (var i = 0; i < spans.length; i++) {
| + | |
| − | spans[i].onclick = function() {
| + | |
| − | for (var index in modals) {
| + | |
| − | if (typeof modals[index].style !== 'undefined') modals[index].style.display = "none";
| + | |
| − | }
| + | |
| − | }
| + | |
| − | }
| + | |
| − | | + | |
| − | // When the user clicks anywhere outside of the modal, close it
| + | |
| − | window.onclick = function(event) {
| + | |
| − | if (event.target.classList.contains('modal')) {
| + | |
| − | for (var index in modals) {
| + | |
| − | if (typeof modals[index].style !== 'undefined') modals[index].style.display = "none";
| + | |
| − | }
| + | |
| − | }
| + | |
| − | }
| + | |
| − | | + | |
| − | </script>
| + | |
| | </body> | | </body> |
| | </html> | | </html> |
| | {{:Team:TUDelft/Footer}} | | {{:Team:TUDelft/Footer}} |
