Yeast transformation (I)

1. Grow 5 ml culture overnight in YPD medium.

2. Take 1 ml and spin down cells. Or take yeast off plate using a toothpick.

3. Resuspend in 0.5 ml PEG/LiAc/TE (40% PEG 4000, 0.1 M Lithium acetate, 10 mM Tris-HCl pH7.5, 10 mM EDTA).

4. Add 10 ml of denatured salmon sperm DNA (10 mg/ml).

5. Add transforming DNA (1-5 mg).

6. Vortex.

7. Let sit on benchtop overnight.

8. Take cells from the bottom layer and plate.