McClean:Mating Type Testers

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Contents

Overview

How to make mating type tester plates from yMM421 and yMM422 and their drug resistant derivatives.

Materials

yMM421/422 colonies growing on YPD YPD and/or Drug plates dH2O Glass Beads

Making the Plates

Each plate will have 100 μL of cells plated on it, so scale up according to the number of plates you need.

  1. Make sure that you use DRY plate, this will speed up the process incredibly.
  2. Dilute 1-2 medium sized single colonies into 500 μL dH2O.
  3. Add 100 uL of this mixture onto each plate.
  4. Add 3-5 glass beads and spread thoroughly.
  5. Allow to dry face up, if they are taking a long time to dry, it is okay to leave the lids slightly ajar under flame.
  6. Once the plates are COMPLETELY dry you can test colonies by pinning or by streaking directly on to the plate.

Assaying Mating Type

  • Make sure to include a mat a, mat α, and diploid control for each plate used to make the the strain is working.
  • Additionally, When using drug resistance markers make sure a WT and known drug resistant control is used.
  • For the assaying drug resistance and mating type simultaneously, make sure to use drug selection plates in a addition to one YPD plate to check viability of the strains on YPD.
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