Team:UC Chile/Labook/march

March

Monthly Summary:

We started with the design of constructs to express RFP under circadian promoters (psbAB and psBA2) and their respective primers. The amplification of the parts needed for the constructs are obtained by PCR. There have been several problems related to the amplification of Kanr. The PCR has been repeated with high frequency with no significant results.

As we decided to do two different projects, the team was divided into subgroups and each person has specific tasks. These are mainly related to dry and wet lab areas.

Constructs and primers design

First constructs. Check: https://2012.igem.org/Team:UC_Chile2/Cyanolux/Biobricks

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.