Team:Linkoping Sweden/Notes

From 2014.igem.org

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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 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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$('#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);
});
});

Revision as of 12:53, 16 October 2014

Here you will find notes of what we did in the lab during this summer and fall.

June

July

August

September

October

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Linköping University
581 83 Linköping, Sweden
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