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From 2011.igem.org

Wednesday

The single A2 colony plated yesterday in pSB1C3 were all red today = no insert!

Transformation of B1 and A2 in pSB1C3 from yesterday. B1 had lot of colonies but red = no insert. A2 had a lot of colonies as well, but some were white! 8 colonies were picked out and replated and an O/N culture made.

Lab Work

• 8 A2 colonies (pSB1C3) were picked out and replated and an O/N culture made.

• Membrane preparation of O/N culture of IPTG induced A1 expressing cells.

• Made overnight cultures for A2 in expression vector (233.3 and 233.4)

• Measured absorption spectra on proteins from the membrane fraction. Despite help from Johan, Thomas, Peter and Birger, we did not succeed to confirm active CYP79A1. There might be a problem with the CO container or the dithionite used (either could be oxidated from the air, and therefore useless).

Other Work

• Had a very nice meeting with Jaide from the DTU-2 Team. She gave us DNA pieces to do User Cloning on CYP79A2 and B1.

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