IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/09/17

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Ligation Procedure:Lumazine/LuxY with plasmid pSB1C3 and promoters J23101/J23102.
I'm working in Step 7 of the Ligation Procedure: Lumazine/LuxY with plasmid pSB1C3 and promoters J23101/J23102.


 * 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 ligated to each promoter, if the ligation was correctly done.

Results:Ligations Lumazine/LuxY with plasmid pSB1C3 and promoters J23101/J23102.Procedure Step 7
After re-culturing the colonies transformed with each ligation mixture, only colonies harboring the Lumazine/LuxY Mr.gene constructions fused to J23101 and J23102 grew correctly.


 * Colonies selected to do the colony PCR reactions:

Lumazine + J23101 ligation from colonies 1,5 and 15.

Lumazine + J23102 ligation from colony 1,6 and 12.

LuxY + J23101 ligation from colony 1, 2 and 15.

LuxY + J23102 ligation from colony 2,4 and 5.




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