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From 2010.igem.org

Precipitation of PCR products

Materials:

- 3 M sodium acetate pH 4.8

- 96% ethanol

- 70% cold ethanol

- MilliQ

- microcentrifuge

- heat block at 37 °C

- nanodrop

Protocol:

1. Add 1/10 volumes of 3 M sodium acetate pH 4.8 to the PCR mixture

2. Add 2 volumes 96% ethanol

3. Incubate 15 minutes at -80 °C or 30 minutes at -80 °C

4. Centrifuge at 14.000 rpm for 10-15 minutes at 4 °C

5. Proceed immediately when centrifuge stops and carefully decant the supernatant

6. Wash DNA pellet with 100 μL of ice cold 70% and centrifuge at 14.000 rpm for 10 minutes at 4 °C

7. Carefully decant the supernatant without disturbing the pellet

8. Air-dry the pellet and redissolve the DNA in water and incubate for 10 minutes at 37 °C

9. Measure DNA concentration on the Nanodrop