Quint Lab:yeast transformation

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yeast transformation - quick and easy

 * 1) 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)
 * 2) Spin down slowly
 * 3) Resuspend in 100 mM LiAc
 * 4) Incubate 45 min (or longer) @30°C
 * 5) Place the volume representing a single 20 . 50 µl inoculum into a separate microcentrifuge tube
 * 6) Add the following components into the tube on top in this order:
 * 7) 240 µl PEG (50% w/v)
 * 8) 36 µl 1.0 M LiAc
 * 9) 50 µl ss-DNA (2.0 mg/ml) (just 5 min boiled before and cooled on ice-water)
 * 10) 5.0 – 30 µl plasmid DNA (100 ng to 5 µg)
 * 11) Vortex 30 s to mix and incubate @ 42°C for 20 min (or longer)
 * 12) Pellet the cells at top speed in a microcentrifuge for 10 – 30 s
 * 13) Remove SN using a micropipet
 * 14) Gently resuspend the pellet in 50 µl of MQ-water by slowly pipetting up and down
 * 15) Plate the cell suspension onto a plate of SC ommision medium that selects for the presence of the plasmid
 * 16) Colonies should be visible in 2 –3 days @ 30°C