|
|
Line 62: |
Line 62: |
| Amplification of parts from 2011 kit: P1003, B0014 and others (specify) are needed for constructs.<br> | | Amplification of parts from 2011 kit: P1003, B0014 and others (specify) are needed for constructs.<br> |
| Standard Biobrick assembly.<br> | | Standard Biobrick assembly.<br> |
- | Transformation.<br>
| + | <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> |
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.