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Imperial College London/Notebook/2 October 2009
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(New page: {{Imperial/09/TemplateTop}} ==Testing== ===OD calibration 4th run 2nd Oct=== ====Protocol considerations==== *Cells were grown overnight in 10ml of M9 media *The cells were allowed to ...) |
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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=== | ||
| + | *18th September (awaiting data analysis) | ||
| + | |||
| + | ===IPTG growth=== | ||
| + | |||
| + | *IPTG has no effect on cell growth under the concentrations tested (100uM to 5mM) | ||
| + | **This is despit the dual effects of possible toxicity and protein production stress by IPTG on Lac-RFP constructs | ||
| + | |||
| + | *16th September (up) | ||
==Testing== | ==Testing== | ||
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*OD of around 0.4 to 1.2 was made up, and subsequently diluted up to 10<sup>-4</sup> to 10<sup>-6</sup> | *OD of around 0.4 to 1.2 was made up, and subsequently diluted up to 10<sup>-4</sup> to 10<sup>-6</sup> | ||
*100ul of cells were inoculated into each plate | *100ul of cells were inoculated into each plate | ||
| - | *The OD of the overnight cells were measured again after 3 hours on ice to check | + | *The OD of the overnight cells were measured again after 3 hours on ice to check if cell number changes during dilution process |
====Results==== | ====Results==== | ||
| - | + | [[Image:II09_OD calib0210graph.jpg]] | |
| - | + | ||
| - | + | ||
| - | |||
| - | + | <br> | |
| + | [[Media:OD-calib02-10.xls| Download raw data]] | ||
| + | |||
| + | ====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==== | ====Suggestions/Improvements==== | ||
Latest revision as of 11:58, 12 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
- 18th September (awaiting data analysis)
IPTG growth
- IPTG has no effect on cell growth under the concentrations tested (100uM to 5mM)
- This is despit the dual effects of possible toxicity and protein production stress by IPTG on Lac-RFP constructs
- 16th September (up)
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
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
