Copenhagen/10 August 2011

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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.
  • Measured absorption spectra on proteins from the membrane fraction. Despite helt 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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