Lissa1:EZ Yeast Transformations
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Preparation of Competent Cells
- Pellet the cells at 500g (setting 3ish on centrifuge) for 4 minutes.
- Discard supernatant.
- Add 10 mL EZ 1 Solution to wash. Repellet and discard supernatant.
- REsuspend in 1 mL EZ 2 Solution
- SLOWLY freeze (in styrofoam container) in -80 freezer.
Transformation
- Mix 50 uL competent cells with .2-1 ug DNA, add 500 uL EZ 3 solution and mix.
- Incubate at 30 C for 45 minutes. Mix by flicking every 15 mins or so.
- Spread 50-150 uL of transformation on your plate.
- Incubate at 30C for 2-4 days to allow for growth of transformants.