Team:BYU Provo/Notebook/SmallPhage/Summerexp/Period2/Exp/7.27 Bacteria Growth Curve After Dilution
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'''I) Purpose''' | '''I) Purpose''' | ||
- | + | * Determine the growth curve of bacteria after dilution. This information will be used to estimate the amount of bacterial growth during incubation in mutagenesis experiments. | |
'''II) Expected Outcome''' | '''II) Expected Outcome''' | ||
- | + | * A sigmoidal growth curve for each dilution. | |
- | + | * The bigger the initial dilution factor, the longer it takes for bacteria to reach its growth plateau. | |
'''III) Reagants Used''' | '''III) Reagants Used''' | ||
- | + | * E. Coli BL21 overnight from 5.26; LB stock; uracil solution (2.5mg/mL); adenine solution (5mg/mL) | |
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1. Bacteria preparation | 1. Bacteria preparation | ||
- | + | * 3mL of BL21 liquid culture overnight was prepared on 7.26. Its OD600 was measured before any dilution (used 2mL). | |
- | + | * Three test tubes were labeled 1:50, 1:100, and 1:500 respectively. These ratios indicate the intended dilution ratios for these test tubes. To each of these three test tubes, 10mL of LB was then added. Each was also supplemented with 40uL of adenine and 80uL of uracil to mimic incubation during mutagenesis | |
- | + | * To the 1:50, 1:100, and 1:500 test tubes, 200uL, 100uL, and 20uL of BL21 liquid culture overnight was then added. | |
2. Measurements using spectrophotometer | 2. Measurements using spectrophotometer | ||
- | + | * Spectrophotometer was warmed up and set to take measurements at 600nm. | |
- | + | * Pure LB was used to as blank | |
- | + | * 1mL of E coli solution at various concentration and time points was pipetted into each cuvette for measurement. | |
- | + | ** The time points include 0, 0.5, 1, 1.5, 2, 2.5, 3.5, 4 hours | |
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1. Raw data | 1. Raw data | ||
+ | |||
+ | OD600 of the BL21 liquid culture overnight is 1.063. | ||
{| class="wikitable" | {| class="wikitable" | ||
|+Data From Spectrophotometer Reading | |+Data From Spectrophotometer Reading | ||
|- | |- | ||
- | | | + | |Time Point |
- | | | + | |1:50 |
- | | | + | |1:100 |
+ | |1:200 | ||
|- | |- | ||
- | |0 | + | |0 h |
- | | | + | |0.037 |
- | |0. | + | |0.0.017 |
+ | |0.001 | ||
|- | |- | ||
- | + | |0.5 h | |
- | |0. | + | |0.179 |
- | |0. | + | |0.132 |
- | + | |0.100 | |
- | + | ||
- | |0. | + | |
- | |0. | + | |
|- | |- | ||
- | | | + | |1 h |
- | |0. | + | |0. |
- | |0. | + | |0. |
+ | |0. | ||
|- | |- | ||
- | | | + | |1.5 h |
- | |0. | + | |0. |
- | |0. | + | |0. |
+ | |0. | ||
|- | |- | ||
- | | | + | |2 h |
- | |0. | + | |0. |
- | |0. | + | |0. |
+ | |0. | ||
|- | |- | ||
- | | | + | |2.5 h |
- | |0. | + | |0. |
- | |0. | + | |0. |
+ | |0. | ||
+ | |- | ||
+ | |3 h | ||
+ | |0. | ||
+ | |0. | ||
+ | |0. | ||
+ | |- | ||
+ | |3.5 h | ||
+ | |0. | ||
+ | |0. | ||
+ | |0. | ||
+ | |- | ||
+ | |4 h | ||
+ | |0. | ||
+ | |0. | ||
+ | |0. | ||
|} | |} | ||
2. Plot | 2. Plot | ||
- | + | ||
3. Linear Regression Result | 3. Linear Regression Result | ||
- | + | ||
'''VI) Conclusion''' | '''VI) Conclusion''' |
Revision as of 01:57, 29 July 2013
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7.27 Bacteria Growth Curve After Dilution
I) Purpose
II) Expected Outcome
III) Reagants Used
1. Bacteria preparation
2. Measurements using spectrophotometer
1. Raw data OD600 of the BL21 liquid culture overnight is 1.063.
2. Plot
3. Linear Regression Result
VI) Conclusion
|