ProtocolsEthanolPrecipitation < Lab

---+ 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*

   1 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.
   1 Add one tenth volume of Sodium Acetate buffer to equalize ion concentrations.
   1 Add at least two volumes of cold 100% ethanol; let stand in -20°C freezer for at least one hour. 
   1 Centrifuge 14,000xg for 30 minutes at 4°C.
   1 Remove as much supernatant as possible with a 1ml micropipet; then remove the rest with a 200ul pipet. 
   1 Add 200ul of cold 70% ethanol; centrifuge for 10 minutes at 4°C. 
   1 Remove supernatant with a 200ul pipet; evaporate remaining ethanol in a 37°C water bath or heat block. 
   1 Resuspend pellet in 50ul of water or TE buffer. 




-- Main.LindseyWolf - 02 Dec 2011

This topic: Lab > WebHome > ProtocolList > ProtocolsEthanolPrecipitation
Topic revision: r2 - 2011-12-05 - LindseyWolf