User:Eduardo Vladimir Munoz/Notebook/UNAM Genomics Mexico 2011/2011/09/13

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High Stabilitiy Plasmid Development Abstract repC-PCR with B0015 ligation colonies replating. repC-PCR with B0015 ligation plasmid extraction. Ligation of P1004 with B0014 Replating I replated four colonies (1-4) that grew after transformation with the ligation of repC-PCR with B0015 in pSB1AK3 in a 6:4 ratio, and ten colonies (5-14) from the ligation with a 7:4 ratio. Plasmid extraction I extracted plasmid using Miguel's protocol from colonies 1, 2, 5, 6 and 9. I will corroborate plasmid concentration tomorrow with an electrophoresis gel. Ligation Since the ratios 7:7 and 9:5 didn't work for the ligation reaction, the ratios 5:9 and 11:3 will be tried: Reaction 5:9 11:3 P1004 5μL 11μL B0014 9μL 3μL T4 DNA ligase 0.5μL 0.5μL 5x ligase buffer 4μL 4μL H2O mQ 1.5μL 1.5μL Total 20μL 20μL