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Protocol: Smash & grab yeast DNA miniprep

  1. Take about 1.5 ml of overnight culture (20-24h) from YPD medium. Spin cells down and remove supernatant.
  2. Add 0.3 g (roughly 0.3 ml) of glass beads, 0.2 ml of lysis buffer and 0.2 ml of a 1:1 mix of phenol and chloroform to the cell pellet in an 1.5 ml tube.
  3. Vortex the tube at top speed for 2 min.
  4. Add 0.2 ml of TE buffer (10 mM Tris, 1 mM EDTA, pH 8.0) and vortex again for a few seconds.
  5. Spin the tubes for 5 min (room temperature) at top speed in an microcentrifuge.
  6. Transfer the aqueous (upper) phase (0.38 ml) to a fresh tube, using a new pipette tip for each sample. Discard the tube with the glass beads.
  7. Add 2 volumes of 100% ethanol at room temperature. Mix thoroughly.
  8. Centrifuge for 2-3 min at room temperature.
  9. Discard the supernatant (use the aspirator; take care not to dislodge the pellet).
  10. Rinse the pellet with 0.5 ml of cold, 70% ethanol, add the ethanol slowly down the side of the tube, then centrifuge for 3-5 sec.
  11. Remove the supernatant. Leave the tubes open and inverted for the pellets to dry (or dry the pellets under vacuum). Resuspend in 50 ┬Ál H2O.


Material:

Glass beads:


Lysis buffer


Source

Based on M. D. Rose, F. Winston, and P. Hieter (1990): Methods in Yeast Genetics: A Laboratory Course Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.