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Team:BCCS-Bristol/Wetlab/0-10mM RFPtest
From 2010.igem.org
(Difference between revisions)
(New page: Having made our double transformants the next step was to characterise their response to Nitrate. We began by examining this between 0 and 10mM of Potassium Nitrate (in 2mM steps). 5mL sol...) |
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Having made our double transformants the next step was to characterise their response to Nitrate. We began by examining this between 0 and 10mM of Potassium Nitrate (in 2mM steps). 5mL solutions of LB and Potassium Nitrate were made up in sterilins and inoculated with the same colony of MG1655s. These were then left overnight at 37°C and their fluorescence analysed using the photo-spectrometer the next morning. This gave results as follows (Where data is missing, colonies produced neither GFP nor RFP, thus were discounted as failures, note how this highlights the usefulness of the double transform method in stopping false negatives): | Having made our double transformants the next step was to characterise their response to Nitrate. We began by examining this between 0 and 10mM of Potassium Nitrate (in 2mM steps). 5mL solutions of LB and Potassium Nitrate were made up in sterilins and inoculated with the same colony of MG1655s. These were then left overnight at 37°C and their fluorescence analysed using the photo-spectrometer the next morning. This gave results as follows (Where data is missing, colonies produced neither GFP nor RFP, thus were discounted as failures, note how this highlights the usefulness of the double transform method in stopping false negatives): | ||
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{| border="1" | {| border="1" | ||
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|- | |- | ||
| 0 | | 0 | ||
| - | | | + | |0.045 |
| - | | | + | |0.0205 |
| - | | | + | |0.45556 |
|- | |- | ||
| 2 | | 2 | ||
| - | | | + | |0.061 |
| - | | | + | |0.1057 |
| - | | | + | |1.73279 |
|- | |- | ||
| 4 | | 4 | ||
| - | | | + | |-- |
| - | | | + | |-- |
| - | | | + | |-- |
|- | |- | ||
| 6 | | 6 | ||
| - | | | + | |0.0522 |
| - | | | + | |0.1128 |
| - | | | + | |2.16092 |
|- | |- | ||
| 8 | | 8 | ||
| - | | | + | |-- |
| - | | | + | |-- |
| - | | | + | |-- |
|- | |- | ||
| 10 | | 10 | ||
| - | | | + | |0.0429 |
| - | | | + | |0.1185 |
| - | | | + | |2.76224 |
|} | |} | ||
| - | |||
| - | |||
| - | |||
Revision as of 10:17, 18 October 2010
Characterising the Ratio Method
Having made our double transformants the next step was to characterise their response to Nitrate. We began by examining this between 0 and 10mM of Potassium Nitrate (in 2mM steps). 5mL solutions of LB and Potassium Nitrate were made up in sterilins and inoculated with the same colony of MG1655s. These were then left overnight at 37°C and their fluorescence analysed using the photo-spectrometer the next morning. This gave results as follows (Where data is missing, colonies produced neither GFP nor RFP, thus were discounted as failures, note how this highlights the usefulness of the double transform method in stopping false negatives):
| Nitrate (mM) | GFP | RFP | GFP / RFP |
| 0 | 0.045 | 0.0205 | 0.45556 |
| 2 | 0.061 | 0.1057 | 1.73279 |
| 4 | |||
| 6 | 0.0522 | 0.1128 | 2.16092 |
| 8 | |||
| 10 | 0.0429 | 0.1185 | 2.76224 |
