Difference between revisions of "Ethanol Precipitation of Vector DNA"
From GcatWiki
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# After digestion with restriction enzymes (ex. Normally a 40 µL digestion) | # After digestion with restriction enzymes (ex. Normally a 40 µL digestion) | ||
| − | # Add | + | # Increase volume to 90 µL with dH20 |
| − | # Add 2X the | + | # Add 10 µL of 3M NaOAc pH 5.2 |
| + | # Add 2X the volume of ethanol (ex. 200 µL of ethanol) | ||
# Vortex and put in -80°C freezer for 15 minutes | # Vortex and put in -80°C freezer for 15 minutes | ||
| − | # Centrifuge for | + | # Centrifuge for 30 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 | + | # Dump off the liquid and allow to dry, either sitting on the bench or with the lyophilizer |
Latest revision as of 14:53, 4 June 2015
- After digestion with restriction enzymes (ex. Normally a 40 µL digestion)
- Increase volume to 90 µL with dH20
- Add 10 µL of 3M NaOAc pH 5.2
- Add 2X the volume of ethanol (ex. 200 µL of ethanol)
- Vortex and put in -80°C freezer for 15 minutes
- Centrifuge for 30 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
