Imperial College London/Notebook/11 September 2009
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+ | {{Imperial/09/TemplateTop}} | ||
+ | ==Testing== | ||
+ | |||
+ | ===Diauxic growth 11th Sep=== | ||
+ | |||
+ | ====Protocol considerations==== | ||
+ | *Using spectrophotometer at 600nm | ||
+ | *Blanks are all the relevant carbon sources, but subsequently realised that all blanks were roughly the same (within 0.03) | ||
+ | **Therefore, subsequent readings= blank + 6 sample | ||
+ | |||
+ | *Overnight culture had OD of 1.64. | ||
+ | **800ul was put into each sample of 40ml~ 0.033 OD | ||
+ | **ie same initial OD | ||
+ | *Grown in 6 separate 50ml falcon tubes and shaking in incubator at 28 degrees Celsius | ||
+ | *1ml taken out at 1 hr intervals from the falcon tube and placed into spectrophotometer | ||
+ | |||
+ | ====Results==== | ||
+ | |||
+ | [[Image:Diauxic growth.jpg]]<br> | ||
+ | [[Media:Diauxic growth_11-09.xls| Download raw data]] | ||
+ | |||
+ | ====Conclusions==== | ||
+ | |||
+ | *Glucose, as expected, had the highest growth rate | ||
+ | |||
+ | *Cells under all secondary carbon source conditions appear to stop growth at 6 hrs. | ||
+ | ** Therefore, we can suggest that the switch point is after 6 hours | ||
+ | **However, further data at longer time points are required to confirm this. | ||
+ | |||
+ | ====Suggestions/Improvements==== | ||
+ | |||
+ | * Longer duration of taking readings (but currently limited to 8hrs- length of a day and multiplate reader evaporation) | ||
+ | |||
+ | * Earlier switch point can be induced by adding a smaller concentration of glucose to media | ||
+ | **suggested 0.01% Glucose for next experiment (we predict it will take around 2 hours) | ||
+ | |||
+ | {{Imperial/09/TemplateBottom}} |
Latest revision as of 16:44, 4 October 2009
Contents |
Testing
Diauxic growth 11th Sep
Protocol considerations
- Using spectrophotometer at 600nm
- Blanks are all the relevant carbon sources, but subsequently realised that all blanks were roughly the same (within 0.03)
- Therefore, subsequent readings= blank + 6 sample
- Overnight culture had OD of 1.64.
- 800ul was put into each sample of 40ml~ 0.033 OD
- ie same initial OD
- Grown in 6 separate 50ml falcon tubes and shaking in incubator at 28 degrees Celsius
- 1ml taken out at 1 hr intervals from the falcon tube and placed into spectrophotometer
Results
Conclusions
- Glucose, as expected, had the highest growth rate
- Cells under all secondary carbon source conditions appear to stop growth at 6 hrs.
- Therefore, we can suggest that the switch point is after 6 hours
- However, further data at longer time points are required to confirm this.
Suggestions/Improvements
- Longer duration of taking readings (but currently limited to 8hrs- length of a day and multiplate reader evaporation)
- Earlier switch point can be induced by adding a smaller concentration of glucose to media
- suggested 0.01% Glucose for next experiment (we predict it will take around 2 hours)