Rather Rapid Genomic Prep
Hoffman and Winston, Gene, 1987
1. Grow 5 ml yeast cultures to saturation.
2. Collect cells by centrifugation and resuspend in 0.5 ml of water. Transfer cells to 1.5 ml microfuge tube and collect by a 5 second spin.
3. Pour off supernatant and briefly vortex to resuspend cells in residual liquid.
4. Add 0.2 ml of Buffer A (2% Triton X-100, 1% SDS, 100 mM NaCl, 10 mM Tris-HCl pH 8.0, 1 mM EDTA pH 8.0), 200 µl glass beads, and 0.2 ml phenol:chloroform:isoamyl alcohol (25:24:1).
5. Vortex 3 minutes (setting #7 on a foam multi-tube vortex adaptor). Add 0.2 ml TE.
6. Spin 5 minutes. Transfer aqueous to new tube. OPTIONAL: Do a chloroform extraction.
7. Add 1 ml 100% EtOH (RT; cold EtOH will create a large, dirty pellet), invert tube to mix, and spin 2 minutes.
8. Discard supernatant, and resuspend pellet in 0.4 ml TE (no need to dry pellet).
9. Add 10 µl 4M ammonium acetate, mix, and then add 1 ml 100% EtOH and mix.
10. Spin for 2 minutes and dry pellet. Resuspend in 50 µl TE and use 10 µl per digest.
2. Collect cells by centrifugation and resuspend in 0.5 ml of water. Transfer cells to 1.5 ml microfuge tube and collect by a 5 second spin.
3. Pour off supernatant and briefly vortex to resuspend cells in residual liquid.
4. Add 0.2 ml of Buffer A (2% Triton X-100, 1% SDS, 100 mM NaCl, 10 mM Tris-HCl pH 8.0, 1 mM EDTA pH 8.0), 200 µl glass beads, and 0.2 ml phenol:chloroform:isoamyl alcohol (25:24:1).
5. Vortex 3 minutes (setting #7 on a foam multi-tube vortex adaptor). Add 0.2 ml TE.
6. Spin 5 minutes. Transfer aqueous to new tube. OPTIONAL: Do a chloroform extraction.
7. Add 1 ml 100% EtOH (RT; cold EtOH will create a large, dirty pellet), invert tube to mix, and spin 2 minutes.
8. Discard supernatant, and resuspend pellet in 0.4 ml TE (no need to dry pellet).
9. Add 10 µl 4M ammonium acetate, mix, and then add 1 ml 100% EtOH and mix.
10. Spin for 2 minutes and dry pellet. Resuspend in 50 µl TE and use 10 µl per digest.
