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1. genDNAprep

1. genDNAprep - Worm Genomic DNA Preparation Reagents...

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Unformatted text preview: Worm Genomic DNA Preparation Reagents Needed: M9 genomic DNA lysis buffer proteinase K phenol/chloroform 3M sodium acetate (common stock) 100% ethanol 70% ethanol ddH 2 O or Buffer TE Procedure: 1. Grow several 10 cm plates of N2 worms on NGM plates overlaid with 1% agarose (agar contains material that can inhibit subsequent enzymatic manipulations of DNA). NOTE: You will probably need to feed the plate at least once to get a lot of worms. Put a few drops of concentrated OP50 on each plate every few days. 2. Wash the worms from the plate with M9 and pellet the worms by spinning at full speed for about 1 minute. 3. Aspirate off supernatant. 4. Add fresh M9, spin, and aspirate supernatant. 5. Repeat step 4 several more times if needed. 6. Flash freeze pellet in liquid nitrogen. ← stopping point 7. Add five volumes of worm genomic DNA lysis buffer with proteinase K (common stock in -20 ° C stock freezer)....
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1. genDNAprep - Worm Genomic DNA Preparation Reagents...

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