Syto RNASelect Green Fluorescent Cell Stain in Yeast — Optimized for quiescent cells after starvation on Day9
- Making working solution
- Bring 5mM stock solution to room temperature
- Quick spin to deposit DMSO solution to bottom of vial
- Make 1mL of 5uM working solution by adding 1uL of stock to 1mL of 1XPBS
- Warm working solution to 37℃ and use immediately (may precipitate if not used).
B. Staining cells
- to a 250uL sample of 5X106 cells add 250uL of stock working solution (final conc. 2.5uM)
- Incubate for 30 minutes at 37℃.
- Quick spin
- Resuspend in 500uL of PBS
C. Fixing (optional)
- Fix in pre-chilled methanol for ten minutes at -20℃.
- Wash 2x in PBS
D. Negative control (optional)
Treat one sample with RnaseA for 1h @ 50℃. Stain samples according to step B.