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Phenol-Chloroform Extraction of DNA (PCI)

This technique can be used to remove or inactivate proteins (such as restriction enzymes or RNAse) from DNA preps.

  • Start with sample volume between 50 and 400 ul in an eppendorf tube
  • Add an equal volume of PCI (25:24:1 Phenol:Chloroform:Isoamyl Alcohol, pH 7.9) to your sample [Take from LOWER phase]
  • Vortex thoroughly
  • Centrifuge 2 min
  • Remove upper phase into new tube, do not disturb interface.
    • Upper phase will form bubble as its volume gets small
  • Add equal vol of CI (24:1, Chloroform:Isoamyl Alcohol)
  • Vortex
  • Centrifuge 2 min
  • Remove upper phase to new tube
  • Perform ethanol precipitation to finish cleaning DNA