Team:UC Chile/Cyano/Labook/april

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<ul>Lab book:<br>
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<li><a href="https://2012.igem.org/Team:UC_Chile/Labook/march">March</a></li><li><a href="https://2012.igem.org/Team:UC_Chile/Cyano/Labook/april">April</a></li><li><a href="https://2012.igem.org/Team:UC_Chile/Cyano/Labook/may">May</a></li><li><a href="https://2012.igem.org/Team:UC_Chile/Cyano/Labook/june">June</a></li><li><a href="https://2012.igem.org/Team:UC_Chile/Cyano/Labook/july">July</a></li><li><a href="https://2012.igem.org/Team:UC_Chile/Cyano/Labook/august">August</a></li><li><a href="https://2012.igem.org/Team:UC_Chile/Cyano/Labook/september">September</a></li><li><a href="https://2012.igem.org/Team:UC_Chile/Cyano/Labook/october">October</a></li>
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<p><font size= "5">April</font></p>
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<font size="4">Monthly Summary</font>
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PCR runs have been defective so we haven’t been able to obtain all the parts needed to assemble our constructs. Nonetheless, parts for C1.1 and C2.1 amplified and a Gibson  assembly was attempted to join them.  The assembly was incorrect according to a restriction enzyme digestion protocol.
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Synechocytis are growing ok.
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<font size="4"><i>Synechocystis PCC6803</i></font>
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Attempted synechocytis DNA extraction according to protocol. There was no pellet and no DNA.
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Synechocystis cultures were observed under acridine dye and turned out to be axenic. (upload photos). Cultures were reinoculated in fresh BG11.
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Started a growth curve for Synechocystis (that was never finished).
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<font size="4">PCR's</font>
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* 17 PCR’s were run. Some parts  amplified correctly and some did not.
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Comments: DNA from synechocystis  as template is not good enough. Primers  form secondary images of these structures)
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<font size="4">Gibson Assembly</font>
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Constructs whose all parts could be obtained by PCR: C1.1 and C2.1. The Gibson assembly technique was used to build the constructs. Transformation followed. 
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Results:  C1.1≈ 7 colonies<br>
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C2.1≈ 5 colonies<br>
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B1 ≈ 1 colony (bacto construct)<br>
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sfGFP (control) ≈ 15, all red (contamination?)<br>
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A DNA extraction and a restriction enzyme protocols were done in order to verify constructs.
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Cut regions:    C1.1 -> psbAB-RFP<br>
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C 2.1 -> psbA2-RFP
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Results: ?? (I believe size was wrong)
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Latest revision as of 19:16, 26 October 2012

Project: Luxilla - Pontificia Universidad Católica de Chile, iGEM 2012