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Copenhagen/10 August 2011
From 2011.igem.org
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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).
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
