Difference between revisions of "UW-Stout/Caffeine"

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== ''Sensitivity to Caffeine'' ==
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== Yeast Sensitivity to Caffeine ==
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== Purpose ==
 
== Purpose ==
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== Materials ==
 
== Materials ==
-10 mM powdered caffeine  
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* 10 mM powdered caffeine  
-100 mM of caffeine solution
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* 100 mM of caffeine solution
-Wild Type Yeast
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* Wild Type Yeast
-Knock out genes
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* Knock out genes
  
 
== Equipment ==
 
== Equipment ==
-Assay Plate
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* Assay Plate
-Micropipettes  
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* Micropipettes  
-1.5 mL test tubes  
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* 1.5 mL test tubes  
-Incubator set at 30 degrees celsius  
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* Incubator set at 30 degrees celsius  
  
 
== General Protocol ==
 
== General Protocol ==
-Find concentrations of caffeine solution needed for experiment
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* Find concentrations of caffeine solution needed for experiment
-Combine caffeine solution with Wild Type yeast (BY4735) in assay plate  
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* Combine caffeine solution with Wild Type yeast (BY4735) in assay plate  
-Incubate at 30 degrees celsius overnight
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* Incubate at 30 degrees celsius overnight
 +
 
 +
 
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* For our experiment we used concentration levels of .2mM, .5mM, 1mM, 2mM, 5mM, 10mM, 20 mM, 50mM, and 100mM.
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* We combined 10uL of each caffeine solution with 90uL of Wild Type Yeast-our control- (BY4735) in the assay tray
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* After analyzing our data we chose to use our .2mM concentration to test on a variety of different yeast genes including:
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  *YG6235W
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  *YJL043W
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  *YCL002C
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  *YMR090W
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  *YCR100C
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  *YPL068C
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  *YEL035C
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  *YFL064C (Our gene)
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== Calibration Experiment ==
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[[File:Calibration experiment.jpg|Calibration Experiment]]
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The data presented is our calibration experiment with wild type yeast combined with various amounts of caffeine concentration. As you can see, the yeast cells were extremely sensitive to the caffeine, and the only clear data was the .2 mM concentration level. We decided to use that concentration point to test on the knock out genes. Looking at the data, the majority of the yeast cells died at .5mM concentration level.
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*Doubling time was 38.0198 minutes

Latest revision as of 07:04, 10 May 2019


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

The data presented is our calibration experiment with wild type yeast combined with various amounts of caffeine concentration. As you can see, the yeast cells were extremely sensitive to the caffeine, and the only clear data was the .2 mM concentration level. We decided to use that concentration point to test on the knock out genes. Looking at the data, the majority of the yeast cells died at .5mM concentration level.

  • Doubling time was 38.0198 minutes