yeast transformation - quick and easy
- Inoculate 2 ml over night culture (selective medium) (OD 0.6 –0.8 is better, maybe dilute and let grow again for few hours in the morning)
- Spin down slowly
- Resuspend in 100 mM LiAc
- Incubate 45 min (or longer) @30°C
- Place the volume representing a single 20 . 50 µl inoculum into a separate microcentrifuge tube
- Add the following components into the tube on top in this order:
- 240 µl PEG (50% w/v)
- 36 µl 1.0 M LiAc
- 50 µl ss-DNA (2.0 mg/ml) (just 5 min boiled before and cooled on ice-water)
- 5.0 – 30 µl plasmid DNA (100 ng to 5 µg)
- Vortex 30 s to mix and incubate @ 42°C for 20 min (or longer)
- Pellet the cells at top speed in a microcentrifuge for 10 – 30 s
- Remove SN using a micropipet
- Gently resuspend the pellet in 50 µl of MQ-water by slowly pipetting up and down
- Plate the cell suspension onto a plate of SC ommision medium that selects for the presence of the plasmid
- Colonies should be visible in 2 –3 days @ 30°C
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