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From 2011.igem.org

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

• 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.