Candida Albicans Lithium Acetate Transformation

Make 50% PEG the day before transformation is to be done.

Grow up a 5ml culture in YPD + uridine
Inoculate 0.1-0.25 ml of the overnight culture into 50 ml YPD + uridine. Incubate at 30^oC for 5 to 7 hours (cells should double at least twice before harvesting).
Harvest cells in a 50 ml conical tube.
Wash with 10 ml sterile water.
Resuspend pellet in 0.5 ml TELiAc.
To two empty ependorf tubes add 5m l of 10mg/ml ssDNA (boil ssDNA before 1^st use).
To one tube add transforming DNA and gently mix (can add up to 80m of a PCR reaction as transforming DNA). Roughly 1-5m g DNA. The other tube is -DNA control.
Aliquot 0.1 ml of cells into the two ependorf tubes.
Incubate at room temperature for 30 min.
Add 0.7ml of PLATE mix to each tube, mix by inversion or trituration.
Incubate at room temp. overnight.
Heat shock cells at 42^oC for 1 hour.
Gently pellet cells in microfuge (3000 rpm, 3 min.)
Suspend pellet in 0.2 ml sterile H₂O and plate 0.1 ml on two selective plates. Only one plate is needed for the -DNA cells.
Incubate plates at 30^oC for 2-3 days.

Solutions