Crime Scene Samples With Gel Electrophoresis

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Comparing Suspect DNA to Crime Scene Samples with Gel Electrophoresis

Christopher Davis

Lab Partners: Jessica Roubert and Aishat Alimi
TA: Tam-Anh Phan

BSC 2010L
Section: 033
November 1, 2014 Materials and Methods
Restriction Enzyme Digest and Preparation of the DNA Samples
Four microcentrifuge tubes were placed in a rack, labeled and numbered, in order to identify the group and the DNA/restriction enzyme that it held. Each of the tubes initially received 10 microliters of reaction buffer. There were two samples of suspect DNA provided along with two restriction enzymes (EcoRI and HindIII). Tubes labeled 1 and 2 received 15 μL of DNA from suspect one while tubes 3 and 4 received 15 μL of DNA from suspect two. Following that, 15 μL of Enzyme 1 (EcoRI) were added to tubes 1 and 3, and 15 μL of Enzyme 2 (HindIII) were added to tubes 2 and 4. (Table 1). The tubes were then gently tapped on the counter to mix the DNA and enzyme solution followed by incubation at 37°C for 45 minutes. After incubation, 5 μL of 10x gel loading dye were added to each of the four tubes of suspect DNA. The tubes were then placed on ice while the gel was under preparation.
Casting, Preparing, and Staining the Gel
To prepare the 0.8% w/v gel, a solution of 50 mL of 1x TBE Buffer was added to a flask containing 0.4g of agarose. The solution was carefully swirled around to mix the contents and then covered with clear plastic wrap to be heated in the microwave for 1 minute. Once the solution
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