McClean:Mating Type Testers
How to make mating type tester plates from yMM421 and yMM422 and their drug resistant derivatives.
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.
- Make sure that you use DRY plate, this will speed up the process incredibly.
- Dilute 1-2 medium sized single colonies into 500 μL dH2O.
- Add 100 uL of this mixture onto each plate.
- Add 3-5 glass beads and spread thoroughly.
- 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.
- 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.