DNA isolation protocols

The following are a few of the many DNA isolation methods available. A quick "dirty" prep is usually sufficient, while some genotyping may work better with highly purified DNA. Determine empirically which protocol works best for your genotyping.

  1. NaOH extraction (quick "dirty" DNA preparation). Reference: Truett GE et al. 2000. Biotechniques 29(1):52-54
    • Cut 2mm of tail and place into an Eppendorf tube or 96-well plate
    • Add 75ul  25mM NaOH / 0.2 mM EDTA
    • Place in thermocycler at 98ºC for 1 hour, then reduce the temperature to 15°C until ready to proceed to the next step
    • Add  75ul of 40 mM Tris HCl (pH 5.5)
    • Centrifuge at 4000rpm for 3 minutes
    • Take an aliquot for PCR (use 2 ul undiluted, or 2 ul of a 1:100 dilution/reaction)
  2. Phenol/chloroform extraction (highly purified DNA preparation)
  3. DNA extraction from blood