- Grow up some fresh yeast on Y(E)PD media. Using an inoculating loop gentle scrape some off of the plate and very gentle spread it around a fresh plate. Let the plate grow at 30C or less until you see white growth(24-48 hours). The fresh plate should be good for transformations for 1-2 weeks if properly stored at 4C.
- Scrape yeast off of plate using inoculation loop(enough to fill the loop) and mix into 100uL of the transformation buffer in a micro tube. Transformation buffer: 100mM LiAc, 40% PEG, 10mM Tris, 1mM EDTA, 0.1mg/mL of single stranded salmon sperm DNA
- Add 0.1 microgram - 1 microgram of plasmid
- Incubate the micro tube at ~42C for 45 minutes. This can be approximated by letting the tube float in water that is warm but not hot from the faucet.
- Incubate the tube at 30C or less for at least 30 minutes and then plate selective media
- Incubate the plate at 30C or less(room temperature is ok) for at least 2 days or until yeast begin to grow