DNA Precipitation protocol

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Revision as of 03:20, 29 September 2009 by Vaishnavi Ananth (Talk | contribs)
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Solutions/reagents:

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

Equipment:

  • Centrifuge
  • Eppendorf tubes

Steps:

  1. Measure out DNA sample into an Eppendorf tube.
    Add 0.1 volume 3M Sodium Acetate solution.
  2. Add 1 µl of Glycogen.
  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.

  7. Dry the pellet in air for 10 - 15 mins.
    Dry until all the liquid is gone.
  8. Option 1: Add water to pellet.
    (or)
    Option 2: Add buffer to pellet.

    Resuspend the pellet by vortexing/by shaking vigorously.

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