Ethanol Precipitation of DNA
互联网
1023
Materials:
3 M sodium acetate pH 5.2 or 5 M ammonium acetate | |
DNA | |
100% ethanol |
- Measure the volume of the DNA sample.
- Adjust the salt concentration by adding 1/10 volume of sodium acetate, pH 5.2, (final concentration of 0.3 M) or an equal volume of 5 M ammonium acetate (final concentration of 2.0-2.5 M). These amounts assume that the DNA is in TE only; if DNA is in a solution containing salt, adjust salt accordingly to achieve the correct final concentration. Mix well.
- Add 2 to 2.5 volumes of cold 100% ethanol (calculated after salt addition). Mix well.
- Place on ice or at -20EC for > 20 minutes.
- Spin a maximum speed in a microfuge 10-15 min.
- Carefully decant supernatant.
- Add 1 ml 70% ethanol. Mix.
- Spin briefly.
- Carefully decant supernatant.
- Air dry or briefly vacuum dry pellet.
- Resuspend pellet in the appropriate volume of TE or water.