Yu:Genomic prep

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  1. Inoculate yeast into 10mL YEPD culture and grow overnight to saturation
  2. Pellet yeast cells 2 minutes at 2k RPM
  3. Discard supernatant and resuspend pellet in 500μL H2O
  4. Transfer yeast suspension to eppendorf tube
  5. Pellet yeast cells 2 minutes at 2k RPM
  6. Resuspend in 200μL Genomic Detergent Lysis Buffer
  7. Add 200μL Phenol:Chloroform:Isoamyl Alcohol (25:24:1)
  8. Add ~300mg glass beads
  9. Vortex 3-4 minutes
  10. Add 200 μL TE8 and spin down for 5 minutes at maximum speed
  11. Transfer the aqueous layer to a new eppendorf tube
  12. Add 1mL of 100% Ethanol, mix by inversion, and spin down for 2 minutes at maximum speed at 4°C
  13. Discard supernatant

    Optional RNAse treatment
    1. Resuspend nucleic acid pellet in 400μL TE8
    2. Add 3μL of pre-boiled RNAse (10 mg/mL)
    3. Incubate 5 minutes at 37°C
    4. Add 10μL of 4M NH4OAc and 1mL 100% Ethanol. Mix by inversion and spin down for 2 minutes at maximum speed at 4°C
    5. Discard supernatant

  14. Air-dry pellet
  15. Resuspend in 50μL of nuclease-free TE8
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