Difference between revisions of "UW Stout/Cycloheximide FA19"

From SGD-Wiki
Jump to: navigation, search
 
Line 41: Line 41:
  
 
This is the data of the protocol and how we found that well two was the best for this experiment
 
This is the data of the protocol and how we found that well two was the best for this experiment
 +
 +
[[Image:First_Protocol_picture.png]]
 +
 +
As you can see from above only four solutions of cycloheximide allowed the yeast to survive those being well 1,2,3, and 12. This data was very important because we needed a solution that didn't fully kill the yeast but disabled it and the well that did that the best was well two. Which disabled the yeast at the optimal percentage.

Latest revision as of 11:00, 3 December 2019

Protocol Finding Correct Dilution of Cycloheximide

Materials List

  • Wild type yeast (control)
  • Cycloheximide
    • .01 grams
  • Sterile H2O
  • 96 well plate
  • Plate Micro Readers
    • 30 Degrees Celsius
  • Our yeast cells

Equipment

  • Micropipettes
  • Incubator set to 30 degrees Celsius
  • New Brunswick TC-7 Tissue Culture Roller Drum
  • Molecular Devices SpectraMax Plus 384 Microplate Reader

Protocol

  1. Make a .001 solution
  2. Label all wells being used
  3. Enter in 50 Microliters of wild type yeast into each well
  4. Then enter in each dilution mixture 1 for each well
  • Add 1 microliter of .001 solution Cycloheximide in 49 microliters H2O
  • Add 5 microliters of solution .001 Cycloheximide in 45 microliters H2O
  • Add 1 microliters of solution .01 Cycloheximide in 49 microliters H2O
  • Add 5 microliters of solution .01 Cycloheximide in 45 microliters H2O
  • Add 10 microliters of solution .01 Cycloheximide in 40 microliters H2O
  • Add 5 microliters of solution 1 Cycloheximide in 45 microlitersH2O
  • Add 10 microliters of solution 1 Cycloheximide in 40 microliters H2O
  • Add 15 microliters of solution 1 Cycloheximide in 35 microliters H2O
  • Add 25 microliters of solution 1 Cycloheximide in 25 microliters H2O
  • Add 35 microliters of solution 1 Cycloheximide in 15 microliters H2O
  • Add 50 microliters of solution 1 Cycloheximide in 0 microliters H2O
  • Control is just 50 microliters water
  1. Incubate in Microplate reader for 6-8hrs at 30 degrees C
  2. Collect data of which yeast cells did not thrive, but did not die, and find the optimal percentage of diluted cycloheximide

The data we found was that well two worked the best for optimal stressed induced to the yeast cells. Add 5 microliters of solution of .001 cycloheximide in 49 microliters sterile H20. with 45 microliters of water and mix that with the 11 yeast strands.

This is the data of the protocol and how we found that well two was the best for this experiment

First Protocol picture.png

As you can see from above only four solutions of cycloheximide allowed the yeast to survive those being well 1,2,3, and 12. This data was very important because we needed a solution that didn't fully kill the yeast but disabled it and the well that did that the best was well two. Which disabled the yeast at the optimal percentage.