IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/09/11
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Ligation Procedure:LovTAP from Mr.Gene to plasmid pSB1C3
I'm working in Step 7 of the Ligation Procedure: LovTAP_Promoters and BBa_K098991 to plasmid pSB1C3.
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, if the ligation was correctly done.
Results:Ligation Procedure Step 7
After re-culturing the colonies transformed with each ligation mixture, only colonies 2, 5, 6 and 10 harboring the LovTAP amplified product from Mr.Gene fused to plasmid pSB1C3 grew correctly.
Thus, these colonies were selected to do the colony PCR reactions.