Grow yeast in 250 ml YEPD until A600 is 0.2-0.8. There are two ways to do this: Inoculate the YEPD with a patch of yeast in the afternoon and grow O/N at room temperature. Inoculate the YEPD with an overnight culture to A600 of ~0.1 and grow at 30°C for ~4hrs. Before starting: boil salmon sperm DNA (10µg/µl) for 5 min an put on ice. prepare TE/LiAc prepare TE/LiAc/PEG Spin at 1800 rpm, 20°C for 5 min. Resuspend in 50ml sterile H2O at RT. Spin at 1800 rpm, 20°C for 5 min. Resuspend in 10ml TE/LiAc at RT. Spin again. Resuspend in TE/LiAc in a total volume of 1 ml per 0.1 OD600. Low competent cells from yeast on a plate

Scrape a patch of cells off a plate. Resuspend in 1 ml TE/LiAc. Spin, resuspend in 50µl TE/LiAc. Add 1µg plasmid, 5µl freshly boiled salmon spermDNA (10µg/µl) and 300 µl TE/LiAc/PEG. Incubate at 30°C for 30 min. Incubate at 42°C for 15 min. Spin, resuspend in 200µl H2O and plate.