From 2007.igem.org

Inoculate 5mL of LB medium with appropriate antibody (e.g. 5 microliter of ampicillin) with a single bacterial colony and incubate at 37 degrees C overnight with vigorous shaking.

Pour 1.5mL of the above culture into an Eppendorf tube. Centrifuge for 1 min. at 10,000g and discard the supernatant by aspiration.

Resuspend the pellet by vortexing in 100 microliters of ice- cold Solution A: 50mM Glucose 10mM EDTA 25mM Tris Cl (pH 8.0)

Add 200 microliters of freshly prepared Solution B: 0.2M NaOH 1% SDS

invert rapidly 4 times. No vortexing.

Add 150 microliters of ice-cold Solution C: 3M potassium 5M acetate (pH 4.8)

vortex for 10 seconds and place on ice for 15 minutes.

Transfer the supernatant to a fresh tube.

Add an equal amount (400 microliters) of phenol/chroloform (200/200 microliters). Vortex briefly and centrifuge at 10,000g for 2 minutes.

Transfer the supernatant to a fresh tube.

Add two volumes (800 microliters) of 100% ethanol. Vortex and place on ice for 20 minutes. Centrifuge at 10,000g for 15 minutes. Discard the supernatant.

Wash by adding 400 microliters of 70% ethanol. Centrifuge for 10 minutes. Discard the supernatant and allow the tube to dry (e.g. vacuum dessicator).

Add 30 microliters of TE buffer with a small amount of RNAase.