BMBMethod:EtOHDNA

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EtOH Precipitation of DNA

This protocol can be used to concentrate DNA, change buffers, or get rid of interfering materials before restriction enzyme digestion, ligation, or whatever.

  • Add 1/10 volume of 3M sodium acetate (pH 5.2) to DNA solution
  • Add 2.5 volumes of 100% ethanol
  • Vortex, store tube at -80°C for 15 min
  • Centrifuge the tube to pellet DNA at 14,000 rpm for 15 min
  • Carefully pour off the supernatant, taking care not to disturb the pellet
  • Add 150ul 70% ethanol to wash the pellet
  • Centrifuge at 14,000 rpm for 5min
  • Discard the supernatant
  • Spin down to collect any residual supernatant, then pipette it off
  • Dry the pellet in air or in SpeedVac
  • DNA pellet can be re-dissolved in ddH2O or TE buffer.
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