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Team:UC Chile/Labook/march
From 2012.igem.org
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<font size="4">Constructs and primers design</font> | <font size="4">Constructs and primers design</font> | ||
| - | First constructs | + | First constructs (link to constructs) |
Primer sequences | Primer sequences | ||
---------- --------------- ----------------- | ---------- --------------- ----------------- | ||
Strategy for integration construct: | Strategy for integration construct: | ||
| - | Part K292003 is not in the kit. It will be assembled from P1003+B0012+B0014 | + | Part K292003 is not in the kit. It will be assembled from P1003+B0012+B0014 |
| + | |||
| + | <font size="4">Getting parts ready</font> | ||
| + | |||
| + | 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. | ||
| + | |||
| + | <font size="4">Work plan</font> | ||
| + | |||
| + | 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. | ||
{{UC_Chilefooter}} | {{UC_Chilefooter}} | ||
Revision as of 20:58, 23 September 2012
March
Constructs and primers design
First constructs (link to constructs)
Primer sequences
---------- --------------- -----------------
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.
