Lissa1:EZ Yeast Transformations

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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.
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