UW-Stout/Cold Shock
Protocol: Cold Shock
Materials
- 8 Knockout Yeast Strains
- Wild Type Yeast Strain (BY4735)
- Pipetter
- Bucket filled with ice
- Stop watch
- 96-well assay plate
- 9, 200 microliter tubes
Purpose
Using cold we will shock each type of yeast to see if it is more or less sensitive to the stress we did on a wild type of yeast labeled BY4735.
Procedure
- Gather all 9 different yeast strains and place in the hood along with 9, 200 microliter tubes.
- Pipette out 100 microliter of each yeast strain placing them each into their own 200 microliter tube. *CHANGING TIP OF PIPETTER IN BETWEEN EACH STRAIN*After each strain is in their own tube, they are now ready to be placed on ice.
- Place each tube on ice for 55 minutes.
- After the 55 minutes, transfer each yeast strain into the 96-well assay plate. Each tube gets its own well, and make sure to change tips in between each strain. The 96-well assay plate is ready to be tested.