Quick Yeast Genomic Prep for PCR
1. Scrape cells off a plate with a pipet tip or toothpick (pin head amount) and resuspend in 500ul H2O
2. Spin for 15 sec @ top speed to harvest cells
3. Resuspend pellet in 250ul of Lysis Buffer (0.1M Tris pH8, 0.25M NaCl, 50mM EDTA, 1% SDS), vortex
4. Add ~500ul of 0.5mm glass beads per sample and shake in Vivadent for 5 sec
5. Spin for 2min @ top speed to reduce foaming
6. Add 250ul of chloroform and shake in Vivadent for 2 sec
7. Spin for 5 min @ top speed to separate phases
8. Transfer ~200ul of upper phase to a new tube, add 500ul 100% EtOH, mix by inverting
9. Spin for 5 min @ top speed
10. Aspirate supernatant and wash pellet once with 70% EtOH
11. Air dry pellet and resuspend in ~300ul H2O
12. Spin DNA sample before setting up PCR to collect insoluble material on the bottom, use 1ul DNA per 10ul PCR reaction
Lysis Buffer: For 50ml:
0.1M Tris pH85ml 1M Tris
0.25M NaCl2.5ml 5M NaCl
50mM EDTA5ml 0.5M EDTA
1% SDS5ml 10% SDS