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PhenolChloEthprecip

DNA Precipitation

Phenol (removes protein)

  1. add equal volume of Phenol (= tris-saturated Phenol-Chloroform-Isoamyethanol)
  2. vortex
  3. spin 2 minutes at 12000 rpm 4C
  4. transfer supernatant to a fresh tube (avoid aspiration of the interlayer or organic phase)

Chloroform (removes phenol)

  1. add equal volume of Chloroform
  2. vortex
  3. spin 2 minutes at 12000 rpm 4C
  4. transfer supernatant to a fresh tube (avoid aspiration of the interlayer or organic phase)

100% Ethanol (precipitates DNA)

  1. add 0.1 volume 3 M sodium acetate
  2. add 2.5 volumes 100 % Ethanol
  3. vortex
  4. precipitate at:
  5. spin 20 minutes at 12000 rpm 4C
  6. carefully pour out / aspirate supernatant (do not lose DNA-pellet)

70% Ethanol (washes out salt)

  1. carefully add 1 mL cold 70% Ethanol (do not vortex)
  2. spin 10 minutes at 12000 rpm 4C
  3. carefully pour out / aspirate supernatant (do not lose DNA-pellet)
  4. air dry 10 minutes at room temperature (do not overdry, because DNA becomes hard to dissolve)
  5. dissolve in: