(for Southerns, PCR, etc.; from Hoffman & Winston, Gene 57, 267-272 [1987])
- Grow 10 ml culture to saturation.
- Harvest by centrifugation.
- Remove supernatant and resuspend cells in 0.5 ml of dH2O.
- Transfer cells to screw-cap micro centrifuge tube.
- Spin 5 seconds in microfuge, decant sup, briefly vortex tube to resuspend pellet in residual liquid.
- Add 0.2 ml of (2% triton X-100, 1% SDS, 100mM NaCl, 10mM Tris-HCl [pH 8.0], 1mM EDTA).
- Add 0.2 ml of phenol:chloroform:isoamyl alcohol (25:24:1).
- Add acid-washed glass beads to the level of the meniscus of the solution.
- Vortex 5 minutes, or longer, until 90% cells are broken.
- Transfer upper, aqueous layer to fresh tube.
- Add 1/10 vol 3M NaOAc, pH 5.2, and 2.5 vol 100% ethanol.
- Wash pellet once with 70% ethanol (-20°C), spin again.