Editing Ethanol Precipitation of Plasmid DNA

Reagents Needed:<br/>• 3 M sodium acetate pH 5.2 or 5 M ammonium acetate<br/>• DNA<br/>• 100% ethanol<br/>Protocol<br/>1. Measure the volume of the DNA sample.<br/>2. Add 1/10 volume of sodium acetate, pH 5.2, (final concentration of 0.3 M)<br/>&nbsp;- These amounts assume that the DNA is in TE only; if DNA is in a<br/>&nbsp;solution containing salt, adjust salt accordingly to achieve<br/>&nbsp;the correct final concentration.<br/>3. Mix well.<br/>4. Add 2 to 2.5 volumes of cold 100% ethanol (calculated after salt addition). (or 0.6 vol of isopropanol)<br/>5. Mix well.<br/>6. Place on ice or at -20 degrees C for >20 minutes.<br/>7. Spin a maximum speed in a microfuge 10-15 min.<br/>8. Carefully decant supernatant.<br/>9. Add 1 ml 70% ethanol. Mix. Spin briefly. Carefully decant supernatant.<br/>10. Air dry or briefly vacuum dry pellet.<br/>11. Resuspend pellet in the appropriate volume of TE or water.&nbsp;
[[Category:Protocols]]