Team:Newcastle/Project/Labwork/OurProtocols/EthanolPrecip

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Ethanol Precipitation

This protocol is designed to remove the ethanol which is likely to be present in a DNA sample once it has been midi-prepped (the ethanol is found in Wash Solution 2).

What you will need

  • 3M Sodium Acetate (pH 5.2)
  • 100% ethanol
  • 70% ethanol
  • Deionised water or buffer (10mM)
  • Ice bucket (with ice)
  • Eppendorf Tubes
  • Centrifuge


Procedure

  1. Measure the DNA sample volume and then calculate 1/10 of this volume. This is the amount of sodium acetate that needs to be added to the sample - pipette this volume of sodium acetate into the DNA sample.


For example if the DNA sample measures 2ml calculate 10% of this volume (which is 0.2ml). Take up 0.2ml of the sodium acetate and add it to the DNA sample - this should make up a total volume of 2.2ml.

  1. Add 100% ethanol to the solution. To work out the amount of 100% ethanol needed, multiply the volume of the DNA sample (plus sodium acetate) by 3.


For example if the DNA sample (plus sodium acetate) makes up a volume of 2.2ml, multiply the 2.2ml by 3, which gives the amount of 6.6ml. Add this volume of ethanol to the DNA (plus sodium acetate) solution, which should now give a total volume of 8.8ml.



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