Ethanol_precipitation
Precipitating out salts from DNA samples
Materials
- 3M Sodium Acetate buffer, pH 5.2 (store at 4°C)
- Cold 100% Ethanol (-20°C)
- Cold 70% Ethanol in sterile dH2O (-20°C)
- DNA sample
- 4°C Microcentrifuge (or normal microcentrifuge in cold room). All centrifuges should be on "soft" (no brake) setting.
Procedure
- Transfer DNA to a container where it fills one fourth the total volume (a 500ul tube should have no more that 125ul of DNA solution, for example.
- Add one tenth volume of Sodium Acetate buffer to equalize ion concentrations.
- Add at least two volumes of cold 100% ethanol; let stand in -20°C freezer for at least one hour.
- Remove as much supernatant as possible with a 1ml micropipet; reventrifuge, then remove the rest with a 200ul pipet.
- Add 200i; of cold 70% ethanol; centrifuge for 5 minutes at 4°C.
- Remove supernatant with a 200ul pipet; evaporate remaining ethanol in a 37°C water bath or heat block.
- Resuspend pellet in 50ul of water or TE buffer.
-- Main.LindseyWolf - 02 Dec 2011
Barrick Lab > ProtocolList > ProtocolsEthanolPrecipitation
Contributors to this topic
LindseyWolf, JeffreyBarrick, GabrielSuarez, VictorLi, AlvaroRodriguez
Topic revision: r1 - 2011-12-03 - 00:03:21 - Main.LindseyWolf