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Imperial College London/Notebook/10 September 2009
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(New page: {{Imperial/09/TemplateTop}} ==Testing== ===Diauxic growth 10th Sep=== ====Protocol considerations==== *Multiwell plate reader **Using script IGEM Abs, taking readings every 30 min *Ov...) |
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*There is an initial spike. This should be a multiplate reader problem | *There is an initial spike. This should be a multiplate reader problem | ||
| - | **Suggestion is to use a higher starting OD ~0.5 or 1 | + | **Suggestion is to use a higher starting OD ~ 0.5 or 1 |
*The diauxic growth is using normal cells which grow on Strep. However, the growth profile might be significantly different on Amp (for our testing construct) | *The diauxic growth is using normal cells which grow on Strep. However, the growth profile might be significantly different on Amp (for our testing construct) | ||
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*Try not to use any readings from the multiplate reader after around 8 hours | *Try not to use any readings from the multiplate reader after around 8 hours | ||
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| + | *To arrive at the switch point faster, 0.01% Glucose will be used instead for next experiment | ||
{{Imperial/09/TemplateBottom}} | {{Imperial/09/TemplateBottom}} | ||
Latest revision as of 20:49, 4 October 2009
Contents |
Testing
Diauxic growth 10th Sep
Protocol considerations
- Multiwell plate reader
- Using script IGEM Abs, taking readings every 30 min
- Overnight culture cells were grown. (normal Top10)
- A volume of overnight cells were diluted into fresh M9 media
- 1ml of each secondary carbon source media (in M9) was prepared
- 200ul was added to 96 well plate as blank
- Write out well plate layout and do test runs of script *IMPORTANT*
- After about 5 hours, when cells were at a high enough OD (cloudy), cells were added 1:20 into the media
- the cells and media were mixed well, and then 200ul was added into 96 well plates
- The plate reader was run overnight
Results
Conclusions
- Cells in Maltose grow slower, while the growth rate in the other secondary carbon sources are similar
- The cells appear to have continuous growth up till 7 hours. However, after that, the results become random. This is probably due to the evaporation problem
Suggestions/Improvements
- There is an initial spike. This should be a multiplate reader problem
- Suggestion is to use a higher starting OD ~ 0.5 or 1
- The diauxic growth is using normal cells which grow on Strep. However, the growth profile might be significantly different on Amp (for our testing construct)
- Therefore, use a cell with an Amp construct that is not expressed
- Try not to use any readings from the multiplate reader after around 8 hours
- To arrive at the switch point faster, 0.01% Glucose will be used instead for next experiment
