Difference between revisions of "UW-Stout/G-418"

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(Created page with "==G-418 Stress== # A In 96-well assay plate in well 1 add 90 ul of yeast YCR100C into the well plate with 10 ul of water for the control. # B In the same well plate in well 2...")
 
(→‎G-418 Stress)
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==G-418 Stress==
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==G-418 Calibration==
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# A In 96-well assay plate in well 1 add 90 ul of yeast YCR100C into the well plate with 10 ul of water for the control.  
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# A In 96-well assay plate in well 1 add 90 ul of yeast BY4735 into the well plate with 10 ul of water for the control.  
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# B In the same well plate in well 2 add 90 ul of yeast YCR100C into the well plate with 10 ul of geneticin with a concentration of 100 ul/ml
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# B In the same well plate in well 2 add 90 ul of yeast BY4735 into the well plate with 10 ul of Geneticin (also known as G-418) with a concentration of 100 ul/ml
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# C In the same well plate in well 3 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 50 ul/ml
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# D In the same well plate in well 4 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 25 ul/ml
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# E In the same well plate in well 5 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 12.5 ul/ml
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# F In the same well plate in well 6 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 6.3 ul/ml
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# G In the same well plate in well 7 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 3.1 ul/ml
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# H In the same well plate in well 8 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 1.56 ul/ml
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# I In the same well plate in well 9 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 0.78 ul/ml
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# J In the same well plate in well 10 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 0.39 ul/ml
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# K In the same well plate in well 11 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 0.2 ul/ml
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# L In the same well plate in well 12 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 0.1 ul/ml
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==Calibration Data==
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From the graphs, we determined that 100 ul/ml was the best option to use for our knock-out experiment because it slowed down the growth of BY4735.
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==Knock-out Experiment==
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# A In 96-well assay plate in well 1 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
 +
# B In same well assay plate in well 2 add 90 ul of yeast YEL035C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
 +
# C In same well assay plate in well 3 add 90 ul of yeast YCL200C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
 +
# D In same well assay plate in well 4 add 90 ul of yeast YFL064C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
 +
# E In same well assay plate in well 5 add 90 ul of yeast YGL235W into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
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# F In same well assay plate in well 6 add 90 ul of yeast YCR100C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
 +
# G In same well assay plate in well 7 add 90 ul of yeast YJL043W into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
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# H In same well assay plate in well 8 add 90 ul of yeast YPL068C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
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# I In same well assay plate in well 9 add 90 ul of yeast YMR090W into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml

Revision as of 07:10, 25 April 2019

G-418 Calibration

  1. A In 96-well assay plate in well 1 add 90 ul of yeast BY4735 into the well plate with 10 ul of water for the control.
  2. B In the same well plate in well 2 add 90 ul of yeast BY4735 into the well plate with 10 ul of Geneticin (also known as G-418) with a concentration of 100 ul/ml
  3. C In the same well plate in well 3 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 50 ul/ml
  4. D In the same well plate in well 4 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 25 ul/ml
  5. E In the same well plate in well 5 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 12.5 ul/ml
  6. F In the same well plate in well 6 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 6.3 ul/ml
  7. G In the same well plate in well 7 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 3.1 ul/ml
  8. H In the same well plate in well 8 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 1.56 ul/ml
  9. I In the same well plate in well 9 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 0.78 ul/ml
  10. J In the same well plate in well 10 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 0.39 ul/ml
  11. K In the same well plate in well 11 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 0.2 ul/ml
  12. L In the same well plate in well 12 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 0.1 ul/ml

Calibration Data

From the graphs, we determined that 100 ul/ml was the best option to use for our knock-out experiment because it slowed down the growth of BY4735.


Knock-out Experiment

  1. A In 96-well assay plate in well 1 add 90 ul of yeast BY4735 into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
  2. B In same well assay plate in well 2 add 90 ul of yeast YEL035C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
  3. C In same well assay plate in well 3 add 90 ul of yeast YCL200C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
  4. D In same well assay plate in well 4 add 90 ul of yeast YFL064C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
  5. E In same well assay plate in well 5 add 90 ul of yeast YGL235W into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
  6. F In same well assay plate in well 6 add 90 ul of yeast YCR100C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
  7. G In same well assay plate in well 7 add 90 ul of yeast YJL043W into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
  8. H In same well assay plate in well 8 add 90 ul of yeast YPL068C into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
  9. I In same well assay plate in well 9 add 90 ul of yeast YMR090W into the well plate with 10 ul of G-418 with a concentration of 100 ul/ml
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