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PCR clean up using 3M sodium acetate and chilled absolute ethanol

Source: KAIMRC, NGHA
Date Added: Sat Jan 02 2010
Date Modified: Sat Jan 02 2010
Abstract: Clean up PCR in 1.5 hrs. with a very easy steps.
Procedure

  1. For each 20 ul of PCR product, prepare the following mixture in 1.5 ml Tube.
    • 2 ul of 3M sodium acetate (NaOAc) pH 4.5
    • 40 ul chilled absolute ethanol.
  2. Transfer the PCR product (20ul) into the tube containing the mixture.
  3. Vortex the tube and then store it at -20 for 30-50 min.
  4. Spin for 30 min at maximum speed 14.000 rpm.
  5. Rremove the supernatant carefully and don't disturb the pellet. ( Do not touch the bottom of the tube)
  6. Wash the pellet with 70% ethanol. And spin the tube at maximum speed for 5 min.
  7. Remove the supernatant and then dry the pellet using vacuum centrifuge. (do not apply heat)
  8. Resuspend the pellet in 30 ul in sterile water.
     
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