Copenhagen/10 August 2011

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===Wednesday===
===Wednesday===
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The single A2 colony plated yesterday in pSB1C3 were all red today = no insert!
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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.
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'''Lab Work'''
'''Lab Work'''
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* 8 A2 colonies (pSB1C3) were picked out and replated and an O/N culture made.
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* Membrane preparation of O/N culture of IPTG induced A1 expressing cells.
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* Made overnight cultures for A2 in expression vector (233.3 and 233.4)
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* 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).   
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[[image:CO Spectra - CYP79A1 - iGEM.jpg|400px|right]]
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'''Other Work'''
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* 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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Back to [[Team:Copenhagen/Notebook|Notebook]]

Latest revision as of 12:55, 11 August 2011

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).
CO Spectra - CYP79A1 - iGEM.jpg

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.


Back to Notebook