1. Harvest 1.5 ml of an ON yeast culture grown under selective conditions (5K, 5 minutes).
2. Resuspend in 100 µl STET (8% sucrose, 50 mM Tris pH 8, 50 mM EDTA, 5% Triton X-100).
3. Add 0.2 g of 0.45 mm glass beads and vortex vigorously for 5 minutes.
4. Add another 100 µl of STET, vortex briefly and place in a boiling water bath for 3 minutes.
5. Cool briefly on ice and spin in a microfuge for 10 minutes at 4°C.
6. Transfer 120 µl of sup to a new tube containing 60 µl of 7.5 M ammonium acetate, incubate 1 hour at -20°C and then centrifuge for 10 minutes at 4°C.
7. Add 120 µl of the sup to 240 µl of ice-cold ethanol, incubate 30 minutes at -20°C and then centrifuge for 10 minutes at 4°C.
8. Wash with 70% ethanol and resuspend in 25 µl TE.
9. Check 5 µl on a gel, use about 10 µl for E. coli transformation

 Good for recovery of plasmids into E.coli.