Minipreps Protocol

Plasmid Minipreps (Alkaline Lysis)

  1. Spin down 1.5ml of culture at 13000 rpm for 1 min
  2. Resuspend the bacterial pellet in 100µl of Solution I
  3. Add 200µl of Solution II. Mix.
  4. Add 150µl of Solution III. Mix.
  5. Spin at 13000 rpm for 1 min.
  6. Transfer the supernatant to a fresh tube and add 400µl of TE Phenol Chloroform.
  7. Vortex, spin at 13000 rpm 1 min.
  8. Transfer the aqueous phase to a fresh tube and add 1ml of Ethanol (100%). Mix.
  9. Spin 1 min at 13000 rpm.
  10. Wash the pellet in 400µl of 70% ethanol.
  11. Vacuum dry the pellet and resuspend in 30µl of TE + 10µg/ml RNAse A.