1) Grow 2ml culture o/n.
2) Spin down. Pour off all but
~100ul of sup. Resuspend cells in the ~100ul.
3) Add 130ul P1. Resuspend.
4) Add 130ul P2. Mix immediately.
5) Add 182ul N3. Mix immediately.
6) Spin 10 min at 4C. (Removes
genomic DNA and debris).
7) Transfer sup to a fresh
eppendorf.
8) Add 900ul ice-cold 100% EtOH
(bottom shelf in door of -20C freezer). Mix thoroughly.
9) Spin 15min at 4C. Discard sup.
10) Wash pellet once with ~400ul
ice-cold 70% EtOH.
11) Dry pellet on rack or in
speedy-vac.
12) Resuspend in 50ul water. Use
3-10ul for digest.
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