Difference between revisions of "PhiX:DNA precipitation"
From CCGB
(Created page with '* 1 volume of isopropanol or 2.5 volumes of 100% ethanol * 0.1 volume 5M ~NaCl (or ~NaAc to 0.3M) * one hour at -80<sup>o</sup>C or O/N at -20<sup>o</sup>C * warm to 0<sup>o</sup…') |
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* 1 volume of isopropanol or 2.5 volumes of 100% ethanol | * 1 volume of isopropanol or 2.5 volumes of 100% ethanol | ||
| − | * 0.1 volume 5M | + | * 0.1 volume 5M NaCl (or NaAc to 0.3M) |
* one hour at -80<sup>o</sup>C or O/N at -20<sup>o</sup>C | * one hour at -80<sup>o</sup>C or O/N at -20<sup>o</sup>C | ||
* warm to 0<sup>o</sup>C (on ice; else viscosity impairs spin) | * warm to 0<sup>o</sup>C (on ice; else viscosity impairs spin) | ||
Revision as of 15:14, 14 August 2009
- 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.
