Yu:Genomic prep
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- Inoculate yeast into 10mL YEPD culture and grow overnight to saturation
- Pellet yeast cells 2 minutes at 2k RPM
- Discard supernatant and resuspend pellet in 500μL H2O
- Transfer yeast suspension to eppendorf tube
- Pellet yeast cells 2 minutes at 2k RPM
- Resuspend in 200μL Genomic Detergent Lysis Buffer
- Add 200μL Phenol:Chloroform:Isoamyl Alcohol (25:24:1)
- Add ~300mg glass beads
- Vortex 3-4 minutes
- Add 200 μL TE8 and spin down for 5 minutes at maximum speed
- Transfer the aqueous layer to a new eppendorf tube
- Add 1mL of 100% Ethanol, mix by inversion, and spin down for 2 minutes at maximum speed at 4°C
- Discard supernatant
Optional RNAse treatment- Resuspend nucleic acid pellet in 400μL TE8
- Add 3μL of pre-boiled RNAse (10 mg/mL)
- Incubate 5 minutes at 37°C
- Add 10μL of 4M NH4OAc and 1mL 100% Ethanol. Mix by inversion and spin down for 2 minutes at maximum speed at 4°C
- Discard supernatant
- Air-dry pellet
- Resuspend in 50μL of nuclease-free TE8