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Copenhagen/26 July 2011
From 2011.igem.org
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*PCR run on the B1 colonies in the expression vector | *PCR run on the B1 colonies in the expression vector | ||
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*Plasmid prep was done on the overnight cultures | *Plasmid prep was done on the overnight cultures | ||
Revision as of 13:59, 26 July 2011
Morning
- The Transformed A2-1 and A2-2 showed no colonies
- The Gel done for yesterdays PCR showed no bands
Lab Work
- A new ligation of A2-1 and A2-2 with PSB1C3 was performed and they were transformed into XL1-BLUE cells
- Digested more PSB1C3
- PCR run on the B1 colonies in the expression vector
- Gel run on the PCR product from B1 colonies
- Plasmid prep was done on the overnight cultures
- Digestion of Plasmid prep with EcoRI and PstI
Other Work
Damian requested the sequence for the RBS we are using. We sent him the sequence of BBa_J04500 (which has the RBS: BBa_B0034).
Since we dont know which plant CYP we will be able to express (if any...) we asked Damian, if he and his team could generate a regulating system that targets two enzymes at the same time. The idea was that the CYPs we use are homologues and have 100 % conserved regions long enough to meet the requirements of their system. In an alignment between A1 and A2, and B1 and A2, two regions of 14 and 9 bases long, repectively, were found. We have asked Damian to use theese sequences in the RNA system.
