Team:Linkoping Sweden/Notes
From 2014.igem.org
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<div class="text-panel"> | <div class="text-panel"> | ||
- | <p> | + | <p>Here you can find notes giving a detailed insight into what we have attempted in the lab during this summer and fall. Read and enter the exciting Lab World of the LiU iGEM Team of 2014! </p> |
</div> | </div> | ||
+ | <div style="width:1100px;display:block;overflow:hidden"> | ||
<div id="june" class="text-panel dropdown"> | <div id="june" class="text-panel dropdown"> | ||
<h2 class="clickable ">June</h2> | <h2 class="clickable ">June</h2> | ||
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</div> | </div> | ||
</div> | </div> | ||
- | + | ||
+ | <div style="width:1100px;display:block;overflow:hidden"> | ||
+ | <div id="august" class="text-panel dropdown"> | ||
+ | <h2 class="clickable ">August</h2> | ||
+ | <div id="augustexp" class="expandable"> | ||
+ | <p>This month we didn’t do much work in the laboratory since we focused more on human practice.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div style="width:1100px;display:block;overflow:hidden"> | ||
+ | <div id="september" class="text-panel dropdown"> | ||
+ | <h2 class="clickable ">September</h2> | ||
+ | <div id="septemberexp" class="expandable"> | ||
+ | <h3>September 8th</h3> | ||
+ | <p>Plasmid preparation on epitope sequence and RFP PSB1C3-E1010.</p> | ||
+ | <p>Restriction digest on epitope, RFP PSB1C3-E1010 and backbone. The epitope was cleaved with EcoRI + SpeI, RFP PSB1C3-J06504 with XbaI + PstI and the backbone with EcoRI + PstI.</p> | ||
+ | <p>Clean-up on the digest products.</p> | ||
+ | |||
+ | <h3>September 9th</h3> | ||
+ | <p>Ligation of epitope sequence, RFP PSB1C3-E1010 and linearized backbone (according to protocol from Thermo Scientific).</p> | ||
+ | <p>Control of digest and ligation on agarosegel.</p> | ||
+ | |||
+ | <h3>September 10th</h3> | ||
+ | <p>PCR with primers on His-TEV.</p> | ||
+ | <p>Clean-up on His-TEV.</p> | ||
+ | <p>Transformation of epitope + RFP (E1010).</p> | ||
+ | |||
+ | <h3>September 11th</h3> | ||
+ | <p>Re-streak of epitope + RFP (E1010).</p> | ||
+ | |||
+ | <h3>September 12th</h3> | ||
+ | <p>Over-night culture of epitope + RFP (E1010).</p> | ||
+ | |||
+ | <h3>September 15th</h3> | ||
+ | <p>Plasmid preparation and glycerol stock on epitope + RFP (E1010).</p> | ||
+ | <p>Digest on His-TEV and RFP backbone (PSB1C3-J06504).</p> | ||
+ | <p>Clean-up on His-TEV and RFP backbone (PSB1C3-J06504).</p> | ||
+ | <p>Gel extraction kit on RFP backbone (PSB1C3-J06504) (to purify the backbone).</p> | ||
+ | <p>Ligation of His-TEV + backbone.</p> | ||
+ | |||
+ | <h3>September 17th</h3> | ||
+ | <p>Transformation of His-TEV + backbone.</p> | ||
+ | <p>Plasmid preparation and glycerol stock of pUC-plasmid.</p> | ||
+ | |||
+ | <h3>September 18th</h3> | ||
+ | <p>Plasmid preparation on RFP backbone (E1010) and RFP backbone (J06504).</p> | ||
+ | <p>Re-streak of His-TEV + backbone.</p> | ||
+ | |||
+ | <h3>September 22nd</h3> | ||
+ | <p>Digest of RFP backbone (J06504) with SpeI and EcoRI.</p> | ||
+ | <p>Clean-up on RFP backbone (J06504).</p> | ||
+ | <p>Gel extraction kit on backbone.</p> | ||
+ | <p>Ligation with purified backbone and His-TEV.</p> | ||
+ | <p>Over-night culture with RFP (J06504).</p> | ||
+ | <p>RFP (J06504) from glycerol stock was put on agar plate with chloramphenicol resistance.</p> | ||
+ | |||
+ | <h3>September 23rd</h3> | ||
+ | <p>Agarose gel with His-TEV after ligation.</p> | ||
+ | <p>Plasmid preparation on RFP (E1010), RFP (J06504) + Epitope and RFP (E1010) + Epitope.</p> | ||
+ | <p>Cultivation on agar plate with RFP (J06504) from glycerol stock.</p> | ||
+ | |||
+ | <h3>September 24th</h3> | ||
+ | <p>Over-night culture with RFP (J06504).</p> | ||
+ | |||
+ | <h3>September 25th</h3> | ||
+ | <p>PCR on RFP (E1010) + epitope and RFP (J06504) + epitope.</p> | ||
+ | <p>Transformation of His-TEV.</p> | ||
+ | |||
+ | <h3>September 29th</h3> | ||
+ | <p>Digest on RFP (E1010) + pUC plasmid with epitope x5.</p> | ||
+ | <p>Digest on RFP (E1010) + pUC plasmid with epitope x5 + backbone.</p> | ||
+ | <p>Digest on RFP (J06504) + pUC plasmid with epitope x5 + backbone.</p> | ||
+ | <p>Clean-up on all three.</p> | ||
+ | <p>Ligation on all three.</p> | ||
+ | <p>Transformation of all three.</p> | ||
+ | |||
+ | <h3>September 30th</h3> | ||
+ | <p>Colony-screening on all three products from September 29th.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div style="width:1100px;display:block;overflow:hidden"> | ||
+ | <div id="october" class="text-panel dropdown"> | ||
+ | <h2 class="clickable ">October</h2> | ||
+ | <div id="octoberexp" class="expandable"> | ||
+ | <h3>October 1st</h3> | ||
+ | <p>Digest on RFP (E1010) + pUC plasmid with epitope 2.</p> | ||
+ | <p>Digest on RFP (E1010) + pUC plasmid with epitope 2 + backbone.</p> | ||
+ | <p>Digest on Digest + pUC plasmid with epitope 2 + backbone.</p> | ||
+ | <p>Clean-up on all three.</p> | ||
+ | <p>Ligation on all three.</p> | ||
+ | <p>PCR-colony screening on all products from the clean-up and ligation.</p> | ||
+ | |||
+ | <h3>October 2nd</h3> | ||
+ | <p>All six products from October 1st was run on agarose gel.</p> | ||
+ | <p>PCR on the His-TEV sequence.</p> | ||
+ | |||
+ | <h3>October 3rd</h3> | ||
+ | <p>New agarose gel for the six products from October 1st since the last one gave bad results.</p> | ||
+ | <p>Digest, clean-up and ligation on His-TEV.</p> | ||
+ | <p>Transformation of His-TEV.</p> | ||
+ | <p>The digest, clean-up and ligation products of His-TEV was run on agarose gel.</p> | ||
+ | |||
+ | <h3>October 4th</h3> | ||
+ | <p>PCR colony screening on His-TEV, RFP (E1010) + epitope x5, RFP (J06504) + epitope x5, RFP (E1010) + epitope 2 and RFP (J06504) + epitope 2.</p> | ||
+ | <p>The PCR samples was loaded on an agarose gel.</p> | ||
+ | <p>Over-night culture on RFP (E1010) + epitope x5, RFP (J06504) + epitope x5, RFP (E1010) + epitope 2 and RFP (J06504) + epitope 2.</p> | ||
+ | <p>Re-streak on the His-TEV transformation from October 3rd.</p> | ||
+ | |||
+ | <h3>October 5th</h3> | ||
+ | <p>Over-night culture on His-TEV.</p> | ||
+ | |||
+ | <h3>October 6th</h3> | ||
+ | <p>Plasmid preparation on on His-TEV, RFP (E1010) + epitope x5, RFP (J06504) + epitope x5, RFP (E1010) + epitope 2 and RFP (J06504) + epitope 2.</p> | ||
+ | |||
+ | <h3>October 8th</h3> | ||
+ | <p>Made starter on RFP (E1010) + epitope 2 and RFP (E1010) + epitope x5.</p> | ||
+ | |||
+ | <h3>October 9th</h3> | ||
+ | <p>Cultivation on RFP (E1010) + epitope 2 and RFP (E1010) + epitope x5.</p> | ||
+ | |||
+ | <h3>October 14th</h3> | ||
+ | <p>FITC of monoclonal and polyclonal antibodies (according to protocol from Thermo Scientific).</p> | ||
+ | |||
+ | <h3>What we’ve planned to do after the wiki-freeze:</h3> | ||
+ | |||
+ | <h3>October 18th</h3> | ||
+ | <p>Cultivation on clones which are correct from sequensation.</p> | ||
+ | |||
+ | <h3>October 19th</h3> | ||
+ | <p>Protein expression on all clones from the cultivation.</p> | ||
+ | |||
+ | <h3>October 20th</h3> | ||
+ | <p>Protein purification.</p> | ||
+ | |||
+ | <h3>October 21st</h3> | ||
+ | <p>Mix of antibodies and expressed proteins (combination of different epitopes and RFP E1010/RFP J06504).</p> | ||
+ | <p>Purify the antibody/protein mixture.</p> | ||
+ | |||
+ | <h3>October 22nd</h3> | ||
+ | <p>Test FRET with fluorescence on all mixtures.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
</div> | </div> | ||
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$('#july').click(function(){ | $('#july').click(function(){ | ||
$('#julyexp').toggle(300); | $('#julyexp').toggle(300); | ||
+ | }); | ||
+ | $('#august').click(function(){ | ||
+ | $('#augustexp').toggle(300); | ||
+ | }); | ||
+ | $('#september').click(function(){ | ||
+ | $('#septemberexp').toggle(300); | ||
+ | }); | ||
+ | $('#october').click(function(){ | ||
+ | $('#octoberexp').toggle(300); | ||
}); | }); | ||
Latest revision as of 20:59, 16 October 2014
Here you can find notes giving a detailed insight into what we have attempted in the lab during this summer and fall. Read and enter the exciting Lab World of the LiU iGEM Team of 2014!