IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/10/02

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Ligation Procedure:Lumazine/LuxY and P0451 with promoter J23102.
I'm working in Step 7 of the Ligation Procedure: Lumazine, LuxY and P0451 with promoter 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 and P0451 ligated to each promoter, if the ligations were correctly done.

Results:Ligations Lumazine, LuxY and P0451 with promoter J23102.Procedure Step 7

 * Colonies selected to do the colony PCR reactions:

P0451 + J23102 ligation from colonies 1,2 and 31.



LuxY + J23102 ligation from colonies 2, 1 and 4.





Lumazine + J23102 ligation from colony 1,5 and 8.



Lumazine + J23102 ligation from colony 2,3,4,6 and 7.



Ligation Procedure: cI inverter(P0451 + K098991) with plasmid pSB1C3. Step 7
I'm working in Step 7 of the Ligation Procedure: cI inverter construction with pSB1C3 backbone.


 * 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 the cI inverter (Parts P0451 + K098991) inside plasmid pSB1C3, if the ligation was correctly done.

Results: Ligation Procedure: cI inverter(P0451 + K098991) with plasmid pSB1C3. Step 7

 * Colonies selected to do the colony PCR reactions:

cI inverter(P0451 + K098991) with plasmid pSB1C3 ligation from colonies 2,3,4,5 and 6.




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