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Imperial College London/Notebook/2 October 2009
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| + | ==Testing team update== | ||
| + | |||
| + | ===Harvard-GFP=== | ||
| + | *There is a difference in fluorescence between 28°C and 37°C after compensating for OD. | ||
| + | *Need to adjust the gain so that fluorescence doesn't saturate | ||
| + | *23rd September | ||
| + | *30th September | ||
| + | |||
| + | ===OD calibration=== | ||
| + | *Preliminary results show that cell number varies with OD. However, the right dilution and enough replicates to overcome the huge variation are required. | ||
| + | *17th of September (up) | ||
| + | *23rd of September (awaiting results) | ||
| + | *25th of September | ||
| + | *2nd October (awaiting results) | ||
| + | |||
| + | ===Secondary carbon sources=== | ||
| + | *There appears to be a plateau in cell number after 6 hours with 0.05% Glucose. However, more data is required to determine if this is the switch point. | ||
| + | *8th September (up) | ||
| + | *10th September (up) | ||
| + | *1st October | ||
| + | |||
| + | ===GFP pH=== | ||
| + | *18September (awaiting data analysis) | ||
| + | |||
| + | |||
| + | ===IPTG growth=== | ||
| + | |||
| + | |||
==Testing== | ==Testing== | ||
Revision as of 20:45, 4 October 2009
Contents |
Testing team update
Harvard-GFP
- There is a difference in fluorescence between 28°C and 37°C after compensating for OD.
- Need to adjust the gain so that fluorescence doesn't saturate
- 23rd September
- 30th September
OD calibration
- Preliminary results show that cell number varies with OD. However, the right dilution and enough replicates to overcome the huge variation are required.
- 17th of September (up)
- 23rd of September (awaiting results)
- 25th of September
- 2nd October (awaiting results)
Secondary carbon sources
- There appears to be a plateau in cell number after 6 hours with 0.05% Glucose. However, more data is required to determine if this is the switch point.
- 8th September (up)
- 10th September (up)
- 1st October
GFP pH
- 18September (awaiting data analysis)
IPTG growth
Testing
OD calibration 4th run 2nd Oct
Protocol considerations
- Cells were grown overnight in 10ml of M9 media
- The cells were allowed to grow until a high OD in the late morning, then placed in cold room
- OD of around 0.4 to 1.2 was made up, and subsequently diluted up to 10-4 to 10-6
- 100ul of cells were inoculated into each plate
- The OD of the overnight cells were measured again after 3 hours on ice to check if cell number changes during dilution process
Results
Waiting to be uploaded
Conclusions
- After 3 hours on ice, the OD stays relatively constant
- Shows that number of cells during OD measurement and during plating stay the same. Therefore, OD correlates with the number of cells that are plated.
Suggestions/Improvements
Waiting to be uploaded
