| Ethanol Precipitation |
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The initial precipitation is done with 100% ethanol (EtOH). This will precipitate DNA as well as the salts that form a Ionic Bond s with it. In a second step, the precipitated pellet is washed with 70%-80% EtOH, which will still precipitate the DNA but remove the salts. BASIC PROTOCOL FOR ETHANOL PRECIPITATION #Add 0.1 volume 3M sodium acetate (NaOAc), pH 5.2 to the DNA to be precipitated. #Add 2.5 volumes 95% ethanol (EtOH), mix gently by inverting. #Incubate 10min on ice. #Centrifuge 20min at max speed on a benchtop microcentrifuge, aspirate supernatant. #Wash once with 1mL 70% EtOH, centrifuge again and aspirate supernatant. #Air-dry 30min and re-suspend in an appropriate volume of Tris-EDTA (TE) |
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