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Draft: Lab Methods 2

Submitted by aspark on Mon, 03/04/2019 - 21:32

The 50 µL of extracted DNA in T10E1 was divided evenly between two tubes, and one was treated with 2 µL of RNase A while the other had 2 µL of sterile water added. Both were incubated for 30 minutes at 37°C before being analyzed by a NanoDrop spectrophotometer. The genomic DNA was also measured via agarose gel electrophoresis. A 50-mL tube of 0.9% agarose containing SYBR Safe was taken from a 65°C water bath, poured into a mold, and set aside to set. A ½ dilution of both the treated and untreated genomic DNA samples were created by combining 10 µL of the DNA stock solutions and 10 µL of dilution buffer. The four loading samples to be run in the gel (genomic DNA treated with RNase, ½ dilution of genomic DNA treated with RNase, untreated genomic DNA, and ½ dilution of untreated genomic DNA) were created by combining 10 µL of each DNA stock solution, 5 µL of loading dye, and 9 µL of water. The gel was loaded with 2.5 µL of DNA ladder in the first lane, 5 µL of DNA ladder in the second lane, and 5 µL of each of the four loading samples in the next four lanes, and the gel was run at 100V for 30 minutes. The gel was visualized via blue light.

 

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