Team:ETH Zurich/labblog/20140702meet

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== Miniprep==
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== Week 6: Project planning, Gibson assemblies planning, First Matlab simulations  ==  
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= Wednesday, July 2nd =
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==== Wednesday, July 2nd ====
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===Miniprep piG0001 piG0029===
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Addition of 200 μL Resuspension solution to culture pellet, vortex, addition of 200 μL Lysis solution <br/>
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After ca. 5 min addition of 350 μL Neutralization solution, centrifuge samples for >10 min at 4000 rpm<br/>
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Preparation of columns: addition of 500 μL Preparation solution, centrifugation for 1 min at 12 000 rcf, discard flow-through<br/>
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Give supernatant of centrifuged samples onto columns, spin for 1 min at 12 000 rcf, discard flow-through<br/>
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Addition of  500 μL Optional Wash Solution, spin for 1 min at 12 000 rcf, discard flow-through<br/>
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Addition of  750 μL Wash Solution, spin for 1 min at 12 000 rcf, discard flow-through<br/>
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Dry columns by centrifuging them  for 2 min at 12 000 rcf, subsequently place columns in new collection tubes<br/>
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Elute DNA with 100 μL Elution solution<br/>
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According to Sigma-Aldrich Plasmid Miniprep Kit protocol<br/>
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Send samples piG0001, piG0015 and piG0029 for sequencing, primers used: oiG0001 for piG0001 and piG0020, oiG0002 for piG0015
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* We have a clear organization of the different experiments we would like to perform.
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[[File:ETH Zurich tasks.PNG|500px]] [[File:ETH Zurich tasks2.PNG|500px]]
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[[#top|Top]]
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We want to optimize quorum sensing to have non-leaky non-cross-talking constructs. We want to prevent cross-talk between integrases and check their dynamics with different quorum sensing promoters. Then we will test the XOR gate without production of LuxI, to check how it works without the loop, and we will finally test our final construct with the loop, hoping that the delay between GFP production and the possible second switching due to AHL production is long enough to have enough fluorescence.
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The red line is our critical timeline. Numbers of days are very optimistic,therefore we multiply the whole timeline by 2. The stars stand for modeling inputs.
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[[File:ETH Zurich tasks3.PNG|center|800px|thumb|Tasks organization with critical timeline in red.]]
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*We have also clear plans for the Gibson assemblies to perform. As an example, here is the plan for regulator plasmids :
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[[File:ETH Zurich regulator progress.png|center|800px|thumb|Exemple of Gibson assembly plan for regulator constructs]]
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* We have a [https://www.facebook.com/iGEM.ETH.Zurich facebook page] !
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[[File:ETH Zurich facebook.PNG|500px|link=https://www.facebook.com/iGEM.ETH.Zurich]]
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* We simulated quorum sensing without leakiness on Matlab, with parameters from the literature.
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<html> </article></html>

Latest revision as of 21:51, 11 October 2014

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