February 15, 2016

DNA Precipitation protocol (2)

Labels:

Solutions/reagents:

  • 3M Sodium Acetate solution
  • Glycogen
  • 95% EtOH
  • 70% EtOH
  • water
  • buffer
  • DNA sample

Equipment:

  • Centrifuge

Steps:

  1. Measure out 0.1 volume 3M Sodium Acetate solution into DNA sample.
  2. Measure out 1 µl of Glycogen into Eppendorf tube (1).
  3. Add 2 volumes 95% EtOH.
  4. Option 1: Store at -20°C for 12 hrs(overnight).
    (or)
    Option 2: Store at -80°C for 30 mins.
  5. Centrifuge at maximum speed for at least 15 mins at room temperature, gently aspirate out the supernatant and discard it.
    6. (Optional)
    Add 1 ml of 70% EtOH.
    Store at room temperature for 5 mins.
    (Optional)
    Centrifuge at maximum speed for 5 mins at room temperature, gently aspirate out the supernatant and discard it.
  6. Dry the pellet in air for 10 - 15 mins.
    Dry until all the liquid is gone.
  7. Option 1: Add water to pellet.
    (or)
    Option 2: Add buffer to water.
    Resuspend pellet by vortexing/by shaking vigorously.
TOTAL TIME REQUIRED FOR THE COMPLETION OF THE PROTOCOL :~ 12 hrs, 25 mins