Lissa1:EZ Yeast Transformations

Preparation of Competent Cells

1. Pellet the cells at 500g (setting 3ish on centrifuge) for 4 minutes.
2. Discard supernatant.
3. Add 10 mL EZ 1 Solution to wash. Repellet and discard supernatant.
4. REsuspend in 1 mL EZ 2 Solution
5. SLOWLY freeze (in styrofoam container) in -80 freezer.

Transformation

1. Mix 50 uL competent cells with .2-1 ug DNA, add 500 uL EZ 3 solution and mix.
2. Incubate at 30 C for 45 minutes. Mix by flicking every 15 mins or so.
3. Spread 50-150 uL of transformation on your plate.
4. Incubate at 30C for 2-4 days to allow for growth of transformants.