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Team:UC Chile/Labook/march
From 2012.igem.org
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Primer alicuoting (primers asked for in late January). | Primer alicuoting (primers asked for in late January). | ||
| - | Amplification of parts from 2011 kit: P1003, B0014 and others (specify) are needed for constructs. | + | Amplification of parts from 2011 kit: P1003, B0014 and others (specify) are needed for constructs.<br> |
| - | Standard Biobrick assembly. | + | Standard Biobrick assembly.<br> |
Transformation.<br> | Transformation.<br> | ||
Plated, only one surviving colony.<br> | Plated, only one surviving colony.<br> | ||
Conclusion: psB1C3 was already CUT<br> | Conclusion: psB1C3 was already CUT<br> | ||
| - | PCR’s. By electrophoresis we have: | + | PCR’s. By electrophoresis we have:<br> |
| - | Unsuccessful attempts: psbAB,psbAB2, Kanr, RS1. | + | Unsuccessful attempts: psbAB,psbAB2, Kanr, RS1.<br> |
| - | Success: RFP, RS2, backbone. | + | Success: RFP, RS2, backbone.<br> |
| - | Comment: seems as primers for expression plasmid are problematic. | + | Comment: seems as primers for expression plasmid are problematic.<br> |
| - | Kanr new PCR attempts. Unsuccessful results. | + | Kanr new PCR attempts. Unsuccessful results.<br> |
| - | PCR for pPMQAK1, psbAB, RFP. | + | PCR for pPMQAK1, psbAB, RFP.<br> |
<font size="4">Work plan</font> | <font size="4">Work plan</font> | ||
Revision as of 21:02, 23 September 2012
March
Constructs and primers design
First constructs (link to constructs)
Primer sequences
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Strategy for integration construct: Part K292003 is not in the kit. It will be assembled from P1003+B0012+B0014
Getting parts ready
Plates prepared Broad host plasmid transformation Batch of new competent bacteria ready Primer alicuoting (primers asked for in late January).
Amplification of parts from 2011 kit: P1003, B0014 and others (specify) are needed for constructs.
Standard Biobrick assembly.
Transformation.
Plated, only one surviving colony.
Conclusion: psB1C3 was already CUT
PCR’s. By electrophoresis we have:
Unsuccessful attempts: psbAB,psbAB2, Kanr, RS1.
Success: RFP, RS2, backbone.
Comment: seems as primers for expression plasmid are problematic.
Kanr new PCR attempts. Unsuccessful results.
PCR for pPMQAK1, psbAB, RFP.
Work plan
Labour division: Bactomithril {Max, Emilia,Ulises, Claudia, Bryon}
Cyano {Simón, Carla, Tamara, Isaac, Sebastián}
Meetings with Dr. Gutiérrez every two weeks to check work done.
