Team:BCCS-Bristol/Wetlab/0-10mM RFPtest

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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
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