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

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1056

 

  • 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.
  • Transfer the PCR product (20ul) into the tube containing the mixture.
  • Vortex the tube and then store it at -20 for 30-50 min.
  • Spin for 30 min at maximum speed 14.000 rpm.
  • Rremove the supernatant carefully and don''t disturb the pellet. ( Do not touch the bottom of the tube)
  • Wash the pellet with 70% ethanol. And spin the tube at maximum speed for 5 min.
  • Remove the supernatant and then dry the pellet using vacuum centrifuge. (do not apply heat)
  • Resuspend the pellet in 30 ul in sterile water.
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