Rapamycin Sensitivity Assay of Yeast
From Bridges Lab Protocols
Revision as of 13:28, 13 October 2009 by Davebridges (Talk | contribs) (Created page with '==Materials== *YPD Media *YPD Plates with either DMSO or 2-200 nM Rapamycin *Yeast strains, streaked out on appropriate plate (< 1 month old) ==Protocol== #Innoculate 3 mL of YP...')
Materials
- YPD Media
- YPD Plates with either DMSO or 2-200 nM Rapamycin
- Yeast strains, streaked out on appropriate plate (< 1 month old)
Protocol
- Innoculate 3 mL of YPD in a small erlenmeyer flask with one colony of yeast
- Grow for ~8h at 24C
- Check OD-600
- Estimate how much to dilute yeast so that the next morning the OD is 0.3. For example, if the doubling time is 2.25h, then from 5 PM to 9 AM is 16h or 7.1 doublings, of 137 times the OD. Therefore to get an OD of 0.3, you need to innoculate at 0.3/137 = 0.00219.
- For each flask innoculate several dilutions
