IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/08/18
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Working on CCaS/CCaR and LuxY synthesized plasmid from Mr.GeneIn order to test the correct lenght for CCaS/CCaR and LuxY synthesized, I am going to do a PCR reaction using the following primers: Forward:Preffix primer Reverse:Suffix primer After the PCR we are going to do a restriction enzyme assay to test the products.
PCRs ResultsAccording with the next image CcaS PCR reaction yields a product around the expected size 3204nt however the fragment amplified in the LuxY PCR reaction has a size around 2000 nt, thus not corresponding to the expected size that is 820 nt. I’m going to repeat the PCR reaction with a shorter Elongation step. Working on LovTAP synthesized plasmid from Mr. GeneLovTAP extraction from gelIn order to extract LovTAP from the gel ready for ligation with J23 promoters and plasmid pSB1C3 , I'm going to do two replicates of enzyme restriction reactions using XbaI and PstI to cut LovTAP plasmid isolated from colony 1, as well I will cut the same plasmid using EcoRI and PstI. LovTAP colony 1.Restriction enzyme assay:Preparation for Ligation with J23 promoters and plasmid pSB1C3
Results:LovTAP colony 1.Preparation for Ligation with J23 promoters and plasmid pSB1C3The plasmid of LovTAP from colony 1 was successfully digested in both restriction enzyme reactions.
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