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Non-technical overview

Mutagenizing a yeast gene with a mini-transposon

First, clone gene into vector pHSS6. Then:

  1. Day 1: Transform plasmid into bacteria expressing transposase.
  2. Day 2: Mate transformants to bacteria carrying mTn (Mating means mixing the cells and plating with selection).
  3. Day 3: Scrape off cells and mate to bacteria carrying resolvase.
  4. Day 4: Scrape off cells, make a miniprep and transform into E. coli.
  5. Day 5: Scrape off transformants, make a miniprep, cut with NotI.
  6. Day 6: Transform digest into yeast.

It's just that simple.

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