PhiX:DNA precipitation

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  • 1 volume of isopropanol or 2.5 volumes of 100% ethanol
  • 0.1 volume 5M NaCl (or NaAc to 0.3M)
  • one hour at -80oC or O/N at -20oC
  • warm to 0oC (on ice; else viscosity impairs spin)
  • centrifuge at full speed for 15 minutes
  • wash pellet in 70% ethanol (this removes salt more efficiently than isopropanol)
  • re-suspend in 10mM Tris, 1mM EDTA, pH 7.5 or in water - may need one hour at 37oC

Remember that isopropanol is a little more efficient (needs less coolth/time to precipitate) but that it precipitates more salt.