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Copenhagen/13 July 2011
From 2011.igem.org
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====Plant CYPs==== | ====Plant CYPs==== | ||
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* We purified A2 (second edition) with [[Team:Copenhagen/Protocol#Mini_prep|Mini prep]] from 2 different colonies. | * We purified A2 (second edition) with [[Team:Copenhagen/Protocol#Mini_prep|Mini prep]] from 2 different colonies. | ||
| - | * We analyzed A2 cut with Pst1 (which cut at the restrictionsite we want to remove)on an agarose gel. Sent them both to be sequences. | + | * We analyzed A2 [[Team:Copenhagen/Protocol#Restrictionsite_analysis|cut]] with Pst1 (which cut at the restrictionsite we want to remove)on an agarose gel. Sent them both to be sequences. |
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'''BioBrick B1''' | '''BioBrick B1''' | ||
| - | * | + | *[[Team:Copenhagen/Protocol#Double Digest|Double digestion]] of B1 using the iGEM approach. |
| + | *[[Team:Copenhagen/Protocol#Ligation Protocol|Ligation]] of the BioBrick (CYP79 B1) and the plasmid backbone (pSB1C3). | ||
| + | *[[Team:Copenhagen/Protocol#Transformation Protocol for pSB1C3|Transformation]] of the constructed plasmid into competent cells. | ||
====Human CYPs==== | ====Human CYPs==== | ||
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Back to [[Team:Copenhagen/Notebook|Notebook]] | Back to [[Team:Copenhagen/Notebook|Notebook]] | ||
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| + | ==Other Work== | ||
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| + | Casper has recorded our CYPerMan song:) | ||
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| + | We have sent the sequences we need to DTU- PowerPuff Team. They will make them into BioBricks that can be assembled with their new assembly method. | ||
Latest revision as of 11:30, 14 July 2011
Contents |
Wednesday
Colonies on A1 - Finally. And our supervisors say that this is the easiest to work with. Hmm.
We still havent heard anything from our nice and helpful benefactors who supplied us with the human CYPs about the sequence and the composition of the plasmid. But we can't wayt any longer so we choose to believe that the plasmid contain ammphicilin. And then well get it sequenced - for this we will use the standard sequence of the middle of the CYPs which has most likely not been altered to much for expression in E.coli.
Lab Work
Plant CYPs
BioBrick A2
- We purified A2 (second edition) with Mini prep from 2 different colonies.
- We analyzed A2 cut with Pst1 (which cut at the restrictionsite we want to remove)on an agarose gel. Sent them both to be sequences.
BioBrick B1
- Double digestion of B1 using the iGEM approach.
- Ligation of the BioBrick (CYP79 B1) and the plasmid backbone (pSB1C3).
- Transformation of the constructed plasmid into competent cells.
Human CYPs
- Transformed XL1-Blue with human cyp 3A4, 2C9, 1A1, 1A2 and hNRP.
Back to Notebook
Other Work
Casper has recorded our CYPerMan song:)
We have sent the sequences we need to DTU- PowerPuff Team. They will make them into BioBricks that can be assembled with their new assembly method.
