FSU - Biological Science

DNA Sequencing Facility - Ammonium Acetate Precipitation

This protocol is useful for cleaning up DNA obtained from mini-prep kits. The procedure is as follows:

  1. Bring volume up to 100 microliters with water.
  2. Add 10 microliters of 5 M Ammonium Acetate and 280 microliters of 95% Ethanol.
  3. Vortex and place at -20 degrees Celsius for 20 minutes.
  4. Spin 5 minutes at room temperature in a micro-centrifuge and remove supernatant.
  5. Add 250 microliters of 70% Ethanol and mix thoroughly.
  6. Spin 5 minutes at room temperature in a micro-centrifuge and remove supernatant.
  7. Dry in speedvac.
  8. Resuspend in water.
  9. Take O.D. to determine concentration.