IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/09/30

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Ligation Procedure:Lumazine/LuxY with plasmid pSB1C3 and promoter J23101.
I'm working in Step 7 of the Ligation Procedure: Lumazine, LuxY and P0451 with plasmid pSB1C3 and promoter J23101.


 * 7.Colony PCR to confirm ligations:

Analyze the colonies with Colony PCR  to confirm that they contain the correct ligation.

The primers that I am using are:

Forward (5'->3'): Preffix primer.

Reverse:(5'->3'): Suffix primer.

These primers would amplify Lumazine, LuxY and P0451 ligated to each promoter, if the ligations were correctly done.

Results:Ligations Lumazine, LuxY and P0451 with plasmid pSB1C3 and promoter J23101.Procedure Step 7

 * Colonies selected to do the colony PCR reactions:

P0451 + J23101 ligation from colonies 1,5 and 23.



LuxY + J23101 ligation from colonies 1, 7 and 21.



LuxY + J23101 ligation from colonies 5,10,11 and 19.





Lumazine + J23101 ligation from colonies 1,4 and 8.



Lumazine + J23101 ligation from colonies 2,3,5,6 and 7.



Lumazine from Mr.Gene + plamid pSB1C3.




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