EtOH / NaOAc Precipitation - Cycle Sequencing


1. Reaction volumes are assumed to be 10 μl.
2. Into each Eppendorf tube add 1 μl of 3M NaOAc, pH 4.6.
3. Into each Eppendorf tube add 25 μl of absolute alcohol (or 95%) – high quality such as from Sigma.
4. Thoroughly mix contents of the tubes.
5. Let precipitate for at least 20 minutes.
6. Centrifuge samples at full speed (14,000 rpm) for 20 minutes, making sure all samples face in the same direction.
7. Remove and discard supernatant.
8. Add 200 μl of 70% alcohol.
9. Centrifuge for 5 minutes at full speed (14,000 rpm), maintaining the same sample direction as before.
10. Remove alcohol, leaving the pellet of DNA behind.
11. Let the DNA pellets air dry.
12. Close tubes, store in cool and dry place until being resuspended and being resolved on a sequencer.

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