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Desalting by Gel Ethanol Precipitation
 
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Desalting Oligonucleotides by Ethanol Precipitation

  1. Measure the volume of the oligonucleotide solution to be precipitated.
  2. Chill 95% Ethanol in the freezer at -20 °C or on dry ice prior to use.
  3. Perform Calculations:

    Volume of 3 Molar Sodium Acetate pH 5.2 = 0.1 * Volume of Oligonucleotide Solution

    Volume of 95% Ethanol = 2.5 * (Volume of Oligonucleotide Solution + Volume of 3 Molar Sodium Acetate pH 5.2)

  4. Add the calculated volume of 3 Molar Sodium Acetate to the oligonucleotide solution and vortex for 10 seconds to mix.
  5. Add the calculated volume of 95% Ethanol to the oligonucleotide solution and vortex for 10 seconds to mix.
  6. Place the reaction mixture in the freezer at -20 °C or on dry ice for at least 1 hour.
  7. Centrifuge the reaction mixture to pellet the precipitated oligonucleotide. A small, gray pellet should now be visible in the bottom of the tube.
  8. Pipette off the supernatant and discard.
  9. Add the same volume of cold 95% Ethanol to the pellet and vortex. It is important to break up the pellet to sufficiently wash out trapped salts.
  10. Repeat Step 7 through 9 once more.

 

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