Team:UC Chile/Cyano/Labook/april

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Latest revision as of 19:16, 26 October 2012

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

April

Monthly Summary

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.

Synechocytis are growing ok.

Synechocystis PCC6803

Attempted synechocytis DNA extraction according to protocol. There was no pellet and no DNA.

Synechocystis cultures were observed under acridine dye and turned out to be axenic. (upload photos). Cultures were reinoculated in fresh BG11.

Started a growth curve for Synechocystis (that was never finished).

PCR's

17 PCR’s were run. Some parts amplified correctly and some did not.

Comments: DNA from synechocystis as template is not good enough. Primers form secondary images of these structures)

Gibson Assembly

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.

Results: C1.1≈ 7 colonies
C2.1≈ 5 colonies
B1 ≈ 1 colony (bacto construct)
sfGFP (control) ≈ 15, all red (contamination?)

A DNA extraction and a restriction enzyme protocols were done in order to verify constructs.

Cut regions: C1.1 -> psbAB-RFP
C 2.1 -> psbA2-RFP

Results: ?? (I believe size was wrong)

@@ Line 31: / Line 31: @@
 <div id="navbar3">
 <ul>Lab book:<br>
-<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/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>
 </ul>
 </div>
@@ Line 41: / Line 41: @@
-[[File:aprillabook_uc_chile.jpg|center]]
+<font size="4">Monthly Summary</font>
+<br><br>
+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.
+Synechocytis are growing ok.
+<font size="4"><i>Synechocystis PCC6803</i></font>
+<br><br>
+Attempted synechocytis DNA extraction according to protocol. There was no pellet and no DNA.
+Synechocystis cultures were observed under acridine dye and turned out to be axenic. (upload photos). Cultures were reinoculated in fresh BG11.
+Started a growth curve for Synechocystis (that was never finished).
+<font size="4">PCR's</font>
+<br>
+<br>
+* 17 PCR’s were run. Some parts  amplified correctly and some did not.
+Comments: DNA from synechocystis  as template is not good enough. Primers  form secondary images of these structures)
+<font size="4">Gibson Assembly</font>
+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.
+Results:  C1.1≈ 7 colonies<br>
+C2.1≈ 5 colonies<br>
+B1 ≈ 1 colony (bacto construct)<br>
+sfGFP (control) ≈ 15, all red (contamination?)<br>
+A DNA extraction and a restriction enzyme protocols were done in order to verify constructs.
+Cut regions:     C1.1 -> psbAB-RFP<br>
+C 2.1 -> psbA2-RFP
+Results: ?? (I believe size was wrong)
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