Team:Linkoping Sweden/Notes

From 2014.igem.org

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

June

July

August

September

October

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Linköping University
581 83 Linköping, Sweden
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