Recombinant DNA - Lab _1 BNS 2.24.11

Recombinant DNA - Lab _1 BNS 2.24.11 - Transfer 700 µL of...

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Supplies/Equipment Cultures in Ice Buckets – 1 per group P1 (stay on ice), water, PE, P2, N3 Vortex Centrifuge (BALANCE) Which pipet to use for each step WHERE is your DNA?? Are you sure?
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Step Protocol: Alkaline Lysis Where is DNA? 1.5 mL of cell culture Centrifuge: 1 min. & remove supernatant 250 µL of Buffer P1: resuspend 250 µL of Buffer P2: invert several times 5 minutes at room temperature Add 350 µL of Buffer N3: invert several times Centrifuge: full speed for 5 minutes
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Unformatted text preview: Transfer 700 µL of supernatant to spin column Centrifuge for 30 sec. & discard the flow-through Add 750 µL of Buffer PE Spin for 30 sec. & discard flow-through Centrifuge: 1 min. to remove residual wash buffer Cut lid off 1.5 mL microfuge tube and place spin column Add 50 µL sterile water to center of column to elute DNA REMINDERS Turn in tube to TA – Label Well Clean-up bench Throw E.coli cultures in WASTE TUBS...
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