IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/09/08
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Ligation Procedure:LovTAP-Promoters and BBa_K098991 to plasmid pSB3K3
I'm working in Step 7 of the Ligation Procedure: LovTAP_Promoters and BBa_K098991 to plasmid pSB3K3.
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 ligated to each promoter and BBa_K098991, if the ligation was correctly done.
Results:Ligations LovTAP-Promoters with plasmid pSB3K3.Procedure Step 7
After re-culturing the colonies transformed with each ligation mixture, only colonies harboring the LovTAP+J23105 (6 and 7) and LovTAP+J23114 (3) fused to plasmid pSB3K3 grew correctly.
Thus, these colonies were selected to do the colony PCR reactions.
Results:Ligation. Part BBa_K098991 with plasmid pSB3K3.Procedure Step 7
After re-culturing the colonies transformed with the ligation mixture, only one colony (1) harboring the Part BBa_K098991(cI regulated promoter+RBS+GFP 933bp) fused to plasmid pSB3K3 grew correctly.
Thus, this colony was selected to do the colony PCR reaction.