Difference between revisions of "UW-Stout/Growth Curve"
(Created page with "==Materials== * [https://www.corning.com/worldwide/en/products/life-sciences/keymatch/3370.html Corning COSTAR 96-well clear flat-bottom assay plate] * [https://www.fishersci....") |
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* [https://www.fishersci.com/shop/products/falcon-round-bottom-polypropylene-tubes-7/p-196968 Falcon round-bottom polypropylene tubes] | * [https://www.fishersci.com/shop/products/falcon-round-bottom-polypropylene-tubes-7/p-196968 Falcon round-bottom polypropylene tubes] | ||
* [https://en.wikipedia.org/wiki/YEPD YPD media], both broth and 2% agar plates | * [https://en.wikipedia.org/wiki/YEPD YPD media], both broth and 2% agar plates | ||
| − | * Glycerol stocks of [UW-Stout/Knockout_Protocol|knockout strains] | + | * Glycerol stocks of [[UW-Stout/Knockout_Protocol|knockout strains]] |
==Equipment== | ==Equipment== | ||
Revision as of 12:15, 29 April 2019
Materials
- Corning COSTAR 96-well clear flat-bottom assay plate
- Falcon round-bottom polypropylene tubes
- YPD media, both broth and 2% agar plates
- Glycerol stocks of knockout strains
Equipment
- Standing incubator set to 30°C.
- New Brunswick TC-7 Tissue Culture Roller Drum
- Molecular Devices SpectraMax Plus 384 Microplate Reader
Protocol
- Three days before the experiment, streak knockout strains onto YPD plates.
- One day before the experiment, pick a colony from each strain into 5 ml YPD broth in a disposable test-tube. Incubate on the roller drum at 30°C overnight.
- The morning of the experiment, dilute each culture to an OD600 of 0.2 in YPD broth. Return to the drum roller for between 30 and 90 minutes.
- Transfer 100 µl to a well in the assay plate.
- Set up the plate reader as follows:
- Temperature: 30°C
- Mode: Kinetic
- Read: 600 nm
- Interval: 5 minutes
- Total run time: 24 hours
- Shake before read: 30 seconds
- Transfer the assay plate to the reader and run for 24 hours.
