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383 lab 3 methods p 4

Submitted by liamharvey on Fri, 04/27/2018 - 12:05

            After selecting primers for our gene, they were ordered from an outside lab source. The primers came back from this lab in 12.5 µL concentrations. Our extracted DNA had to be diluted first by adding ~5 µL of extracted DNA to ~45 µL of T10E1. However, Mutant B was found to be of very low concentration when run in the Nanodrop. For Mutant B only, no T10E1 was used to dilute the DNA sample and 2 µL of DNA was used alone instead. For Wild Type A, which had an initial concentration of 532.3 ng/µL, 5 µL of DNA was added to 45 µL of T10E1. For Wild Type B, with an initial concertation of 360.7 ng/µL, 7.5 µL of DNA was added to 42.5 µL of T10E1. For Mutant A, an initial concentration of 619.1 ng/µL, 4 µL of DNA was added to 46 µL of T10E1. With this exception, we calculated the diluted concentrations of each sample; Wild Type A had a diluted concentration of 53.23 ng/µL, Wild Type B 54.11 ng/µL, Mutant A 49.53, and Mutant B 2 µL undiluted at 6.5 ng/µL.