tRNA, from Brewer's Yeast, 500

Yeast IF without Dehydration

  Grow cells at the appropriate temperature to 5x10E6 in 5 mls YPD. Add 0.5mls 37 formaldehyde (best grade) and incubate on the roller at same temp. for 10 min. Spin down cells 2Kx3min. and resuspend in 5mls 40mMKPO4 pH6.5/500uM

Dropout plates for yeast

to mix together and then add to agar to make plates. Making amino acid dropout (CSM) solution Generally easiest to make a 10x master solution that you can dilute back when you''re pouring plates. Let''s assume you''re going to make 200ml

Yeast Nuclei Isolation

Alternative: Instead of recombinant lyticase, can also use Zymolyase 100T from ICN. Use ~2 mg per 2 liters of cells. The amount required can vary from 2-10 mg depending on the yeast strain. If using zymolyase, add in one extra YPD/S washing step