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Imperial College London/Notebook/17 September 2009
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(New page: ==Testing team== ===OD calibration=== *Use dilutions of 10<sup>-6</sup> to 10<sup>-8</sup> *Problems: **Was not done on ice, so cell growth still occurred during dilutions **Too many colon...) |
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| - | ==Testing | + | {{Imperial/09/TemplateTop}} |
| - | ===OD calibration=== | + | |
| - | * | + | ==Testing== |
| - | * | + | |
| - | **Was not done on ice, so cell growth still occurred during dilutions | + | ===OD calibration 17th Sep=== |
| - | + | ||
| - | + | ====Protocol considerations==== | |
| + | |||
| + | *Cells were grown overnight in 10ml of M9 media | ||
| + | *The cells were allowed to grow until a high OD (very cloudy) and retrieved in the early afternoon | ||
| + | *The OD was measured using the multiplate reader. | ||
| + | *As the OD was only around 1.5, the cells were centrifuged and resuspended in less media to form an OD of 3 | ||
| + | *OD of around 0.5 to 3.0 was made up, and subsequently diluted up to 10<sup>-6</sup> to 10<sup>-8</sup> | ||
| + | *Plated on strep plates and dried until the end of the day | ||
| + | *Placed into 37 degrees incubator (non-shaking) overnight | ||
| + | |||
| + | ====Results==== | ||
| + | |||
| + | The plates had too many colonies to count. | ||
| + | <!--[[Image:Diauxic growth2.jpg]]<br> | ||
| + | [[Media:Diauxic growth_10-09.xls| Download raw data]]--> | ||
| + | |||
| + | ====Conclusions==== | ||
| + | |||
| + | * The plates were all too confluent, even at 10<sup>-8</sup> dilution. | ||
| + | * Unable to count colonies and draw any colonies | ||
| + | * Need to repeat experiment | ||
| + | |||
| + | |||
| + | ====Suggestions/Improvements==== | ||
| + | |||
| + | *Was not done on ice, so cell growth still occurred during dilutions | ||
| + | *Too many colonies to count when plated 100uL | ||
**Use ice | **Use ice | ||
**Increase dilutions | **Increase dilutions | ||
| + | |||
| + | |||
| + | {{Imperial/09/TemplateBottom}} | ||
Latest revision as of 18:13, 4 October 2009
Contents |
Testing
OD calibration 17th Sep
Protocol considerations
- Cells were grown overnight in 10ml of M9 media
- The cells were allowed to grow until a high OD (very cloudy) and retrieved in the early afternoon
- The OD was measured using the multiplate reader.
- As the OD was only around 1.5, the cells were centrifuged and resuspended in less media to form an OD of 3
- OD of around 0.5 to 3.0 was made up, and subsequently diluted up to 10-6 to 10-8
- Plated on strep plates and dried until the end of the day
- Placed into 37 degrees incubator (non-shaking) overnight
Results
The plates had too many colonies to count.
Conclusions
- The plates were all too confluent, even at 10-8 dilution.
- Unable to count colonies and draw any colonies
- Need to repeat experiment
Suggestions/Improvements
- Was not done on ice, so cell growth still occurred during dilutions
- Too many colonies to count when plated 100uL
- Use ice
- Increase dilutions
