IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/09/29

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LovTAP fused to promoters and Lumazine:Ligations to backbone pSB3K3 and pSB1C3
I'm working in Step 7 of the Repeated experiment:Working on LovTAP fused to promoters and Lumazine:Ligations to backbone pSB3K3 and pSB1C3.


 * 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 LovTAP fused to promoters and Lumazine, if the ligations were correctly done.

Results:LovTAP fused to promoters and Lumazine:Ligations to backbones pSB3K3 and pSB1C3.Procedure Step 7

 * Colonies selected to do the colony PCR reactions:

Ligations to plasmid pSB3K3

LovTAP + J23117 isolated from colonies 2, 3 and 4.

LovTAP + J23105 isolated from colonies 1, 2 and 3.

LovTAP + J23114 isolated from colonies 9,13 and 14.

LovTAP + J23102 isolated from colonies 1,11 and 13.

Ligations to plasmid pSB1C3

LovTAP + J23117 isolated from colonies 1,6 and 7.

LovTAP + J23105 isolated from colonies 1, 12 and 13.

LovTAP + J23114 isolated from colonies 1,2 and 3.

LovTAP + J23102 isolated from colonies 1,2 and 8.

Lumazine (Mr.Gene) colonies 1 and 2.








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