Team:TU Dresden/Notebook/Protocols/DNAPrecipitation


DNA precipitation

  1. Transfer the DNA into a 1.5 mL tube.
  2. Add 0.1 Vol. of 3 M NaAc to it.
  3. Add 2.5 Vol. of 100 % EtOH or at least 0.6 Vol. 2-propanol and mix.
  4. Incubate at RT for 20 minutes; incubation at -20 °C may improve yield.
  5. Spin down the DNA at 13,200 rpm for 10 minutes.
  6. Discard the ethanol, but be careful not to lose the DNA-pellet.
  7. Carefully add 150 μL 70 % EtOH to the pellet, centrifuge again at 13,200 rpm for 5 min if the pellet is only loosely attached to the wall of the tube.
  8. Either decant or pipette out the 70 % EtOH with the large pipette, then short spin and then use a pipette with a yellow tip to remove the remaining 70 % EtOH, but be careful not to lose the DNA-pellet .
  9. Dry the pellet for 5-10 min at room temperature, or place the opened tube in a heating block at 50 °C. Be careful to not over dry the pellet.
  10. Re-dissolve the pellet in 12 μL of water, TE–buffer or 10 mM Tris/HCl pH 8.5.