Quint Lab:yeast plasmid prep

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yeast plasmid prep

  1. inoculate 2 ml of Glucose medium and let grow over night @ 30°C
  2. spin down 1.5 ml of the over night culture and resuspend in0.5 ml S-buffer
  3. incubate 30 min, 37°C
  4. add 0.1 ml lysis solution, vortex
  5. incubate 30 min, 65°C
  6. add 0.166 ml potassium acetate and mix, chill on ice for 10 min
  7. spin down for 10 min, pour the SN into a new tube (2 ml!)
  8. DNA precipitation: add 0.8 ml EtOH, 10 min on ice, spin down for 10 min, pour off the SN
  9. wash DNA with 0.5 ml 70% EtOH, dry pellet
  10. dissolve DNA in 0.04 ml water


S-buffer:

  • 10 mM K2HPO4; pH 7.2
  • 10 mM EDTA
  • 50 mM 2-Mercaptoethanol
  • 50 µg/ml zymolase


lysis solution:

  • 25 mM Tris-HCl; pH 7.5
  • 25 mM EDTA
  • 2.5% SDS
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