Team:Paris Saclay/ethanol

From 2013.igem.org

Ethanol precipitation

This method is used to purify and/or concentrate DNA, RNA and polysaccarides.

Protocol :

1. Measure the volume of the DNA sample.

2. Add 3x volumes of ethanol 100%

3. Add 1/10x volume of sodium acetate (3 M, pH 5.2)

4. Add 1µL of glycogen

5. Incubate the solution at -20°C for 30min or more

6. Centrifuge at > 14,000 x g for 30min at 4°C

7. Discard supernatant being careful not to throw out DNA pellet which may or may not be visible.

8. Wash with 1mL of ethanol 70%

9. Centrifuge at > 14,000 x g for 15min at 4°C

10. Discard supernatant, air dry pellet and dissolve pellet in desired buffer or water