Difference between revisions of "Ethanol Precipitation of Vector DNA"
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− | + | # After digestion with restriction enzymes (ex. Normally a 40 µL digestion) | |
− | + | # Add 1/10 the digestion volume of 3M NaOAc pH 5.2 (ex. 4 µL of NaOAc) | |
− | + | # Add 2X the reaction volume of ethanol (ex. 88 µL of ethanol) | |
− | + | # Vortex and put in -80°C freezer for 15 minutes | |
− | + | # Centrifuge for 15 minutes (Place the tube hinge out in the microcentrifuge to know where the pellet is. | |
− | + | # Dump off the liquid and allow to dry, either sitting on the bench or with the lyophilizer. |
Revision as of 13:15, 30 June 2011
- After digestion with restriction enzymes (ex. Normally a 40 µL digestion)
- Add 1/10 the digestion volume of 3M NaOAc pH 5.2 (ex. 4 µL of NaOAc)
- Add 2X the reaction volume of ethanol (ex. 88 µL of ethanol)
- Vortex and put in -80°C freezer for 15 minutes
- Centrifuge for 15 minutes (Place the tube hinge out in the microcentrifuge to know where the pellet is.
- Dump off the liquid and allow to dry, either sitting on the bench or with the lyophilizer.