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Method: Salting Out Procedure for Human DNA Extraction

April 24, 1990

C. Helms


Time required:

Special solutions:


Day 1

  1. Isolate nuclei from 1-2 tubes of blood (collected in ACD or EDTA tubes):

    Add 9 volumes Buffer A, mix well and hold on ice 2 minutes. Spin at 1500 rpm at 4deg.C for 15 minutes. Resuspend the nuclei pellet in 5 ml Buffer B and transfer to a 15 ml polypropylene centrifuge tube.

  2. Add 500 ul 10% SDS and 55 ul proteinase K (10 mg/ml stock). Incubate at 37deg.C overnight on a low-speed rocker or orbital shaker.
Day 2

  1. Add 1.4 ml saturated NaCl soution (approximately 6M) to each tube. Shake vigorously for 15 seconds. Spin the tubes at 2500 rpm in the Beckman low speed centrifuge for 15 minutes.
  2. Transfer the supernatant to another 15 ml polypropylene tube, leaving behind the precipitated protein pellet.
  3. Add exactly two volumes of room temperature 100% ethanol and invert the tube several times until the DNA precipitate is visible.
  4. Remove the DNA strands with a plastic spatula or pipet tip and transfer to an eppendorf tube containing 100 - 200 ul TE. Allow the DNA to dissolve at least 2 hours at 37deg.C before quantitating.



S. A. Miller, Dykes, D. D., and H. F. Polesky . (1988). " A simple salting out procedure for extracting DNA from human nucleated cells" Nucleic Acids Research 16: 1215.