Team:HKUST/Protocols/Ethanol precipitation

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(New page: 8. Ethanol precipitation Purpose: To concentrate DNA product. Materials: 100% ethanol, 75% ethanol, 3M NaAc, ddH2O, DNA product Procedure: a.Add 2 volume of 100% ethanol. b....)
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Revision as of 13:59, 26 August 2009

8. Ethanol precipitation 

  Purpose: To concentrate DNA product.
  Materials: 100% ethanol, 75% ethanol, 3M NaAc, ddH2O, DNA product
  Procedure:
  a.Add 2 volume of 100% ethanol.
  b.Add 0.1 volume of 3M NaAc.
  c.Put the tube in fridge at -20 °C for 10 minutes.
  d.Put it out and centrifuge it for 10-15 minutes (13,000 rpm). Remove the supernatant.
  e.Wash with 100μL 75% ethanol.
  f.Centrifuge for 5 minutes (13,000 rpm). Remove all the supernatant.
  g.Add 10μL ddH2O to resuspend DNA.
  Tips:
  •If the volume of DNA product is too low, make it to higher volume with ddH2O. The recommended lowest volume is 50μL. Combine several tubes of DNA product is also suggested for higher DNA product concentration.
  •In step f, make sure to remove all the supernatant without touching the pellet. If it is too hard to do so, open the tube and leave it at room temperature for a while to make the ethanol evaporate.
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