Phenol-Chloroform purification of DNA

  1. Add an equal volume of phenol-chloroform to sample.
  2. Mix by shaking tube.
  3. Spin 1' in microfuge tube at max speed.
  4. Remove upper (aqueous) layer, avoiding the interface and add to a fresh tube.
  5. Mix with an equal volume of chloroform.
  6. Spin 1' in microfuge, tube at max speed.
  7. Remove upper layer again, add to a fresh tube.

Isoproponal precipitate DNA:

  1. Add 1% volume of 3M Potassium acetate, pH 5.5.
  2. Add 0.7 volumes isopropanol.
  3. Mix, and spin precipitated DNA in microfuge at room temperature at max speed for 10m.
  4. Carefully remove supernatant.
  5. Add 500 μl 70% ethanol to tube.
  6. Spin 5m in microfuge at max speed.
  7. Carefully remove supernatant with vacuum.
  8. Let tubes dry (open) for a few minutes.
  9. Resuspend in your buffer of choice.
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