UW-Stout/Caffeine

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Yeast Sensitivity to Caffeine

Purpose

The purpose of this lab is to test the sensitivity to the caffeine of knock out strains of yeast cells. We will be analyzing the data on the gene's response to different levels of caffeine concentration.


Materials

  • 10 mM powdered caffeine
  • 100 mM of caffeine solution
  • Wild Type Yeast
  • Knock out genes

Equipment

  • Assay Plate
  • Micropipettes
  • 1.5 mL test tubes
  • Incubator set at 30 degrees celsius

General Protocol

  • Find concentrations of caffeine solution needed for experiment
  • Combine caffeine solution with Wild Type yeast (BY4735) in assay plate
  • Incubate at 30 degrees celsius overnight


  • For our experiment we used concentration levels of .2mM, .5mM, 1mM, 2mM, 5mM, 10mM, 20 mM, 50mM, and 100mM.
  • We combined 10uL of each caffeine solution with 90uL of Wild Type Yeast-our control- (BY4735) in the assay tray
  • After analyzing our data we chose to use our .2mM concentration to test on a variety of different yeast genes including:
  *YG6235W 
  *YJL043W
  *YCL002C
  *YMR090W
  *YCR100C
  *YPL068C
  *YEL035C
  *YFL064C (Our gene)

Calibration Experiment

Calibration Experiment