Difference between revisions of "UW-Stout/Co Culture SP21"
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==Equipment== | ==Equipment== | ||
| − | * | + | *[http://www.southwestscience.com/SBT1500H_Heated_Plate_Vortexer.html Southwest Science heated plate shaker set to 600 rpm and 30°C] |
*flow cytometer | *flow cytometer | ||
*type 1000, 200, and 20 micro pipettors | *type 1000, 200, and 20 micro pipettors | ||
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==Protocol== | ==Protocol== | ||
Revision as of 13:51, 13 April 2021
This page is under construction
Materials
- Double strength synthetic-defined media (liquid)
- Single-strength synthetic-defined media (liquid)
- Phosphate buffered saline, sterile.
- Glycerol stocks of knockout strains
- Thomas Scientific square,deep 96 well, round bottom micro assay plate
Equipment
- Southwest Science heated plate shaker set to 600 rpm and 30°C
- flow cytometer
- type 1000, 200, and 20 micro pipettors
Protocol
- Create concentration of 100 cells per micro liter for both the knockout and control fluorescent strain with the Hemocytometer within 1 mL of double strength synthetic media.
- Place contents into a 2 mL culture within a deep well microassay plate.
- Repeat steps 1 and 2 for each additional yeast strain, assigning each a unique well within the microassay plate.
- Incubate the plate within the incubator shaker for 48 hours at 30 degrees Celsius, shaking at 600 rpm.
- From each well, take a 5 micro liter sample and mix it with 500 micro liters of phosphate buffered saline to measure on the flowcytometer.
