1. Grow 1 liter of yeast cells in YPD to stationary phase.
2. Pellet and wash
2x and resuspend in zymolyase buffer.
3. Zymolyase treat
for 1 hour, check for speroblasting
4. Gently pellet
spheroblasts and resuspend in 12.5 mls of lysis buffer
50 mM Tris pH 8.0
50 mM EDTA pH 8.0
1 % Sarkosyl
5. Add 800 ul of
10 mg/ml RNAse A solution, 1 h at 37°C
6. Add 1 ml of
40/mg/ml protienase K 1 h at 37°C
7. CHCl3 extract
8. Add CsCl to
1.71 g/ml and spin.
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