Ethanol Precipitation of Vector DNA

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  1. After digestion with restriction enzymes (ex. Normally a 40 µL digestion)
  2. Increase volume to 90 µL with dH20
  3. Add 10 µL of 3M NaOAc pH 5.2
  4. Add 2X the volume of ethanol (ex. 200 µL of ethanol)
  5. Vortex and put in -80°C freezer for 15 minutes
  6. Centrifuge for 30 minutes (Place the tube hinge out in the microcentrifuge to know where the pellet is)
  7. Dump off the liquid and allow to dry, either sitting on the bench or with the lyophilizer