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.