Hi I have soil DNA extracted via Soil DNA extraction kit, but after running on Gel I couldn't get any fragment. Now I want to precipitate it with Ethanol prior to amplification . Final suspension was n 300 ul of TE Buffer. Kindly tell me how to perform this precipitation ? Thanks
Hello, this is what works for me:
Add 1/10th volume of 3M sodium acetate ph 5.2, and 2X volume cold 100% EtOH. Vortex for 10 seconds and incubate at -80 C for at least 30 min. Spin at 12,000 xg for 30 min and discard supernatant. Wash pellet with cold 70-75% EtOH and spin for 10 min. Discard supernatant and let pellet air dry for 10 min and dissolve in 50ul buffer or DNase/RNase free water.
Well Thanks Newly xyz and Beatriz Kowalski for your good suggestions. Beatriz I have used your mentioned precipitation method and it worked , although DNA concentration was too low, but still it was visible on GEL. Regards
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