Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test27settembre
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*<partinfo>BBa_K173000</partinfo> (positive control) | *<partinfo>BBa_K173000</partinfo> (positive control) | ||
*<partinfo>BBa_B0031</partinfo> (negative control) | *<partinfo>BBa_B0031</partinfo> (negative control) | ||
- | + | Cultures were grown ON at 37°C, 220 rpm. | |
- | The following day cultures were diluted 1:100 and let grow again for about five hours at | + | The following day cultures were diluted 1:100 and let grow again for about five hours at 37°C, 220 rpm. |
- | + | The optical density (O.D.) of each cell culture was than measured with TECAN Infinte F200. Samples were diluted in order to obtain the same O.D. equal to 0,02. | |
- | + | Then we performed a 21-hours' experiment with measurements of absorbance and green fluorescence every five minutes using TECAN Infinite F200; cultures were shaken for 15 seconds every five minutes. Each value shown below is the mean of three measurements, from GFP data that of a non-fluorescent culture (negative control) was subtracted. | |
==RESULTS== | ==RESULTS== | ||
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[[Image:UNIPV_Pavia_SScellM27settembre.png|thumb|500px |center|Mean of (dGFP/dt)/O.D.600 (under the hypothesis that GFP half-life in fusion contructs is similar to the original one - <partinfo>BBa_E0040</partinfo>)]] | [[Image:UNIPV_Pavia_SScellM27settembre.png|thumb|500px |center|Mean of (dGFP/dt)/O.D.600 (under the hypothesis that GFP half-life in fusion contructs is similar to the original one - <partinfo>BBa_E0040</partinfo>)]] | ||
- | All cell cultures showed a similar growth curve | + | All cell cultures showed a similar growth curve; doubling time was computed as described [[Team:UNIPV-Pavia/Parts/Characterization#Doubling_time_evaluation|here]] in order to have informations about the burden due to synthesis of such fusion proteins. It's possible to see that all doubling time are very similar; it's possible to assert that the expression of these BioBrick parts doesn't cause abnormal stress to cells. |
From GFP curve it's possible to appreciate that in I47, I48, I49 GFP accumulation it's very similar and it's significantly different from that of negative control <partinfo>BBa_B0031</partinfo>. This result that the green fluorescent protein assembled downstream of the genetic circuit is correctly folded. | From GFP curve it's possible to appreciate that in I47, I48, I49 GFP accumulation it's very similar and it's significantly different from that of negative control <partinfo>BBa_B0031</partinfo>. This result that the green fluorescent protein assembled downstream of the genetic circuit is correctly folded. |
Latest revision as of 22:24, 27 October 2010
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