Difference between revisions of "UW-Stout/Ultraviolet Light"

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(Materials)
(Materials)
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* [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]]
* [https://ecatalog.corning.com/life-sciences/b2c/US/en/Surfaces/Advanced-Cell-Culture-Surfaces/Corning%C2%AE-Treated-Culture-Dishes/p/430166 Cell 60 mm Culture Dish]
+
* [https://ecatalog.corning.com/life-sciences/b2c/US/en/Surfaces/Advanced-Cell-Culture-Surfaces/Corning%C2%AE-Treated-Culture-Dishes/p/430166 Cell 60 mm Culture Dish, containing 6 ml of agar]
  
 
==Equipment==
 
==Equipment==

Revision as of 06:40, 2 May 2019

Materials

Equipment

  • Incubator set to 30°C.
  • Bachur & Associates Sanata Clara, CA 95050 Model LS-100-3 UV Light Exposeure System

Protocol

  1. Three days before the experiment, streak knockout strains onto YPD plates.
  2. 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.
  3. 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.
  4. Transfer 33 µl to a well in the assay plate.
  5. Set up the UV light Exposeure System as follows:
    • 400 watts
    • time increments
    • place in the incubator in dark for 48 hours

Result