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Purifying DNA

Standard Protocols

Purifying DNA


If at A260/A280 the purity of DNA is out of the 1.8-2.0 range then the DNA should be purified to remove contaminants.

  1. In the fume hood wearing appropriate protection i.e. gloves etc.. Add an equal Volume of Phenol/Chloroform/isopropanol mixture (in fridge) to the DNA. (make sure the tubes used are resistant to phenol before you begin)
  2. Vortex the sample thoroughly, solution should be milky.
  3. Centrifuge at 3,000rpm for 10 minutes.
  4. Transfer the top layer to a clean tube, be careful not to take over the white fluffy layer. If a small amount is carried over then recentrifage at 3,000rpm for 10 minutes and remove the supernatant to a clean tube.
  5. If there is a lot of white fluffy material at the interphase then repeat procedure on the sample once more.
  6. Ethanol precipitate the sample.

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