optimizing glucose concentrations of bacterial cultures
TRANSCRIPT
Optimizing Glucose Concentrations of Bacterial Cultures using E. Coli HB101
An experiment by: Jacob Shercliffe & Ronald Wagoner
Research Question: What is the optimal amount of glucose to grow the most concentrated E.coli HB101 in an LB broth liquid culture?
Hypothesis and Predictions
� Hypothesis to be tested: As the concentration of glucose in the LB broth increases, the concentration of E.coli HB101 that grows will also increase, until it plateaus and then drops off.
� Expected Results: We predicted that since the glucose adds extra food to the culture it would increase the overall concentration, but we also predicted that too much would Choke them out, and possibly raise the pH too much
Methods � Inoculate stock culture with pGLO transformed E.coli HB101.
� Make 0-25% glucose Amp/LB broth in 5% glucose increments.
� Fill culture tubes & inoculate them with pGLO transformed E.coli HB101.
� Perform an OD600 reading each day using a spectrophotometer. to determine the concentration of cells in the culture.
� Get pH readings at the beginning and end of the culturing process.
0
200,000,000
400,000,000
600,000,000
800,000,000
1,000,000,000
1,200,000,000
1,400,000,000
1,600,000,000
0 1 1 2 2 3 3 4 4 5
Conc
entr
atio
ns (
mg/
ml)
Time (Days)
Concentrations of E.coli HB101
0% 5% 10% 15% 20% 25%
Optimizing Glucose Concentrations of Bacterial Cultures using E. Coli HB101!
Bacterial Growth is a common practice in many laboratory experiments that require stock solutions of a bacterium. In this experiment we isolated the growing conditions to find optimal growth through glucose
variation. !
Jacob Shecliffe & Ronald Wagoner, Chugiak High School!
Results:!Discussion:!
Graphic
Graph outlining the growing patterns of the different
concentrations of glucose!
Graphic
Bar graphing highlighting the different pH’s of the samples on
different days!
Introduction:! Methods:!
Graphic
Graphic
A spectrophotometer used for measuring the concentrations !
Graphic
pH probe to measure the acidity of a solution!
– "What is the optimal amount of glucose to grow the most concentrated E.coli HB101 in an LB broth liquid culture?"– "E.coli is a common bacterial strand and HB101 is a glow in the dark lab-safe strain for use in many different experiments" – "This study gives a brief snapshot of how varying the amount of glucose within a liquid culture changes the maximum concentration of bacteria and their growth patterns."
v From our data we conclude that adding some glucose to your liquid culture is much better than adding none"
v We also found that the optimal concentration was between 10 & 15%"
u From the initial pH of 7, all of the samples went down in acidity, especially in the presence of any glucose, then only incrementally based on concentration."
u Glucose created much more growth in every concentration, and differed based on concentration "
• Inoculate stock culture with pGLO transformed E.coli HB101 & make 0-25% glucose Amp/LB broth in 5% glucose increments.
• Perform an OD600 reading each day using a spectrophotometer to determine the concentration of cells in the culture.
• Get pH readings at the beginning and end of the culturing process.
0 1 2 3 4 5 6 7 8
Day 1
Day 3
pH (1-14)
Tim
e (D
ays)
pH of Samples
25%
20%
15%
10%
5%
0%
0
200,000,000
400,000,000
600,000,000
800,000,000
1,000,000,000
1,200,000,000
1,400,000,000
1,600,000,000
0 1 1 2 2 3 3 4 4 5
Conc
entr
atio
ns (
mg/
ml)
Time (Days)
Concentrations of E.coli HB101
0%
5%
10%
15%
20%
25%
0 1 2 3 4 5 6 7 8
Day 1
Day 3
pH (1-14)
Tim
e (D
ays)
pH of Samples
25% 20% 15% 10% 5% 0%