Winston Prep for Isolation of Genomic Yeast DNA
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  1. Grow 5 ml of yeast culture to saturation (1 or 2 days) in YPD (rich media)
  2. Collect cells using a falcon tube in a clinical centrifuge at 3/4 speed around 5 min.
  3. Pour off supernatant and resuspend cells in 500 ul water and transfer to a microfuge tube. Spin in microfuge 10 seconds full speed and pour off water. Use residual water left behind to resuspend the pellet.
  4. To pellets add 200 ul Winston juice, around .3 g (2 bead spoonfuls) acid washed glass beads, and 200 ul phenol/chloroform.
  5. Vortex at full speed 5 minutes.
  6. Add 200 ul TE (10mM Tris, 1mM EDTA) to the preps and microfuge on high 5 min.
  7. Transfer supernatant to new tubes, add 200 ul phenol/chloroform, and vortex again for 3 minutes. Spin for a few sec.
  8. Transfer supernatant to new tube, add 1 ml 100% EtOH, invert to mix, and microfuge on high 5 min. Discard supernatant and last remnants of EtOH. Let pellets air dry a few min.
  9. Resuspend in 400 ul TE, add about 20 ug RnaseA, and incubate at 37°C for 5 minutes. (The pellet is hard to resuspend so don't work too hard. It's much easier after the RnaseA digestion).

Winston Solution
2% Triton X-100
1% SDS
100mM NaCl
10mM Tris pH8

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