IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/09/08: Difference between revisions
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|style="background-color: #EEE"|[[Image: | |style="background-color: #EEE"|[[Image:TeamLogo.jpg|350px]]<span style="font-size:19px;"> UNAM-Genomics-Mexico team</span> | ||
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Revision as of 14:23, 9 October 2010
 UNAM-Genomics-Mexico team
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Ligation Procedure:LovTAP-Promoters and BBa_K098991 to plasmid pSB3K3I'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 7After 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 7After 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. 
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