UW-Stout/Glycerol FA22 - Revision history

Teagueb at 20:39, 13 December 2022

2022-12-13T20:39:11Z

Teagueb at 20:38, 13 December 2022

2022-12-13T20:38:58Z

Teagueb at 20:38, 13 December 2022

2022-12-13T20:38:10Z

Teagueb at 20:37, 13 December 2022

2022-12-13T20:37:44Z

Teagueb at 20:37, 13 December 2022

2022-12-13T20:37:25Z

Teagueb at 20:37, 13 December 2022

2022-12-13T20:37:04Z

Teagueb at 20:27, 13 December 2022

2022-12-13T20:27:39Z

Teagueb at 20:13, 13 December 2022

2022-12-13T20:13:21Z

Teagueb at 20:13, 13 December 2022

2022-12-13T20:13:02Z

Teagueb at 20:12, 13 December 2022

2022-12-13T20:12:03Z

@@ Line 33: / Line 33: @@
 [[File:Yeast_Growth_in_Glucose_Media_Exp._1.png|600px]]
-This graph represents the yeast growth in glycerol media, plotted with the OD600 reading versus time every 5 minutes. A doubling time of around 549 minutes was calculated.
+This graph below represents the yeast growth in glycerol media, plotted with the OD600 reading versus time every 5 minutes. A doubling time of around 549 minutes was calculated.
 [[File:Yeast_Growth_in_Glycerol_Media_Exp._1.png|600px]]

@@ Line 30: / Line 30: @@
 ==Wild Type Yeast Data==
 For our pilot experiment, we had two different samples: wild type yeast grown in 1x glucose media and wild type yeast grown in the 3% glycerol media.
 [[File:Yeast_Growth_in_Glucose_Media_Exp._1.png|600px]]
-The OD600 reading of the yeast cells per time every 5 minutes is graphed on a scatterplot. The growth peaks above 0.5. and a doubling time around 231 minutes was calculated.
+This graph represents the yeast growth in glycerol media, plotted with the OD600 reading versus time every 5 minutes. A doubling time of around 549 minutes was calculated.
 ==Knockout Protocol==
 Based on our plots, it is clear that the yeast in the glucose media grew at a much quicker rate. The glycerol media very obviously stunted the growth of the yeast, but not too much that the yeast died off. It doubled at about half the rate the glucose yeast did. This amount of stress seems like a reasonable amount to test on the knockout strains of yeast, so for the knockout protocol the same procedure will be used on 5 different knockout strains.

@@ Line 31: / Line 31: @@
 For our pilot experiment, we had two different samples: wild type yeast grown in 1x glucose media and wild type yeast grown in the 3% glycerol media.
 [[File:Yeast_Growth_in_Glucose_Media_Exp._1.png|600px]]
 The OD600 reading of the yeast cells per time every 5 minutes is graphed on a scatterplot. The growth peaks above 0.5. and a doubling time around 231 minutes was calculated.
 [[File:Yeast_Growth_in_Glycerol_Media_Exp._1.png|600px]]
 This graph represents the yeast growth in glycerol media, plotted with the OD600 reading versus time every 5 minutes. A doubling time of around 549 minutes was calculated.

@@ Line 30: / Line 30: @@
 ==Wild Type Yeast Data==
 For our pilot experiment, we had two different samples: wild type yeast grown in 1x glucose media and wild type yeast grown in the 3% glycerol media.
-[[File:Yeast_Growth_in_Glucose_Media_Exp._1.png|600px]] The OD600 reading of the yeast cells per time every 5 minutes is graphed on a scatterplot. The growth peaks above 0.5. and a doubling time around 231 minutes was calculated.
+[[File:Yeast_Growth_in_Glucose_Media_Exp._1.png|600px]]
-[[File:Yeast_Growth_in_Glycerol_Media_Exp._1.png|600px]] This graph represents the yeast growth in glycerol media, plotted with the OD600 reading versus time every 5 minutes. A doubling time of around 549 minutes was calculated.
+The OD600 reading of the yeast cells per time every 5 minutes is graphed on a scatterplot. The growth peaks above 0.5. and a doubling time around 231 minutes was calculated.
 ==Knockout Protocol==
 Based on our plots, it is clear that the yeast in the glucose media grew at a much quicker rate. The glycerol media very obviously stunted the growth of the yeast, but not too much that the yeast died off. It doubled at about half the rate the glucose yeast did. This amount of stress seems like a reasonable amount to test on the knockout strains of yeast, so for the knockout protocol the same procedure will be used on 5 different knockout strains.

@@ Line 31: / Line 31: @@
 For our pilot experiment, we had two different samples: wild type yeast grown in 1x glucose media and wild type yeast grown in the 3% glycerol media.
 [[File:Yeast_Growth_in_Glucose_Media_Exp._1.png|600px]] The OD600 reading of the yeast cells per time every 5 minutes is graphed on a scatterplot. The growth peaks above 0.5. and a doubling time around 231 minutes was calculated.
-[[File:Yeast_Growth_in_Glycerol_Media_Exp._1.png]] This graph represents the yeast growth in glycerol media, plotted with the OD600 reading versus time every 5 minutes. A doubling time of around 549 minutes was calculated.
+[[File:Yeast_Growth_in_Glycerol_Media_Exp._1.png|600px]] This graph represents the yeast growth in glycerol media, plotted with the OD600 reading versus time every 5 minutes. A doubling time of around 549 minutes was calculated.
 ==Knockout Protocol==
 Based on our plots, it is clear that the yeast in the glucose media grew at a much quicker rate. The glycerol media very obviously stunted the growth of the yeast, but not too much that the yeast died off. It doubled at about half the rate the glucose yeast did. This amount of stress seems like a reasonable amount to test on the knockout strains of yeast, so for the knockout protocol the same procedure will be used on 5 different knockout strains.

@@ Line 1: / Line 1: @@
 ==Introduction==
-In yeast cells, glucose is a preferred carbon source for fermentation in yeast cells. Fermentation produces ethanol and CO2 from glucose. Other carbon sources, like glycerol, are unfermentable which, in the absence of glucose, would use other forms of carbon fixation with alternate products. In this experiment, we will be testing the results of glucose starvation with a replacement of glycerol as a carbon source in a number of knockout strains of yeast.
+In yeast cells, glucose is a preferred carbon source for fermentation in yeast cells. Fermentation produces ethanol and CO2 from glucose. Other carbon sources, like glycerol, are unfermentable which, in the absence of glucose, would use other forms of carbon fixation. In this experiment, we will be testing the results of glucose starvation with a replacement of glycerol as a carbon source in a number of knockout strains of yeast.
 ==Materials/Equipment==
@@ Line 31: / Line 31: @@
 For our pilot experiment, we had two different samples: wild type yeast grown in 1x glucose media and wild type yeast grown in the 3% glycerol media.
 [[File:Yeast_Growth_in_Glucose_Media_Exp._1.png|600px]] The OD600 reading of the yeast cells per time every 5 minutes is graphed on a scatterplot. The growth peaks above 0.5. and a doubling time around 231 minutes was calculated.

@@ Line 24: / Line 24: @@
 * 100x Lysine Solution
 * 100x Uracil Solution
- Modify the liquid yeast media by replacing the original glucose solution with the glycerol media solution made in step 1. Repeat this for the wild type yeast as well as the 5 knockout strains.
+Modify the liquid yeast media by replacing the original glucose solution with the glycerol media solution made in step 1. Repeat this for the wild type yeast as well as the 5 knockout strains.
 Set up the yeast wells, working underneath the biochemical safety hood, including the controls of each knockout strain.
 Set up the plate reader and run for 18 hours

@@ Line 3: / Line 3: @@
 ==Materials/Equipment==
-*96-well assay plate
+* 96-well assay plate
--Microplate reader
+* Microplate reader
--Wild-type yeast cells
+* Wild-type yeast cells
--5 yeast knockout strains
+* 5 yeast knockout strains
--3% glycerol media (procedure included later in this study)
+* 3% glycerol media (procedure included later in this study)
--Sterile water
+* Sterile water
--P200 and P1000 micropipette and tips
+* P200 and P1000 micropipette and tips
 ==Safety Note==
@@ Line 16: / Line 16: @@
 ==Protocol==
 Make a 3% glycerol media solution using appropriate amounts of materials below:
--Yeast nitrogen base
+* Yeast nitrogen base
--De-ionized water
+* De-ionized water
--Ammonium sulfate
+* Ammonium sulfate
--Pure glycerol
+* Pure glycerol
--Complete supplement mixture
+* Complete supplement mixture
--100x Histone Solution
+* 100x Histone Solution
--100x Lysine Solution
+* 100x Lysine Solution
--100x Uracil Solution
+* 100x Uracil Solution
   Modify the liquid yeast media by replacing the original glucose solution with the glycerol media solution made in step 1. Repeat this for the wild type yeast as well as the 5 knockout strains.
 Set up the yeast wells, working underneath the biochemical safety hood, including the controls of each knockout strain.