UW-Stout/Nitrogen Starvation FA21
Contents
Nitrogen Starvation Experiment
Introduction
The purpose of this experiment is to test different variations of yeast and how they adapt to the pressure of starving them of nitrogen which is considered necessary for there growth and how the yeast react alterations in media. And if any strains of the yeast react better or worse to starvation and if they are able to grow efficiently without a nitrogen source available.
Protocol
Materials/equipment
- 15 plates of media that contains ammonium (nitrogen source)
- 15 plates of media that does not contain ammonium (no nitrogen source)
- 96 well plate
- Micro-pipettors
- Specialized multi-channel micro-pipette
- Micro pipette tips
- 6 PCR tubes containing wild yeast and 5 knockout strains
- 10-15ul of each yeast strain
- Sterile Water
- YNB
- CSM
- His
- Leu
- Ura
- Agar
- H2O
- Ammonium Source
Procedure for Media Formulation
- On two separate containers, two solutions of media with 1/2 L each will be made. One with Nitrogen and one without.
- add required amount of water followed by all required ingredients in descending order from materials section.
- Place media solutions in Steam Sterilizer Machine for about 40 minutes.
- plate the media onto required amount of plates evenly spread.
Experimental Procedure
- Obtain all necessary materials and make sure equipment is functional and ready for use.
- Obtain 2 media plates for each yeast strain, 1 plate with ammonium and 1 plate without ammonium for each strain.
- label all media plates and PCR tubes
