Team:Aachen/Interlab Study
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We introduced the three constructs in to ''E. coli'' cells and measured fluorescence as well as optical density of the liquid cultures over a period of 18 hours, using a spectrophotometer and a plate reader, respectively. The obtained results confirmed our hypothesis that the fluorescence of the BioBrick in the high copy plasmid pSB1C3, J23101.E0240, would exhibit a stronger signal than the constructs I20260, which is on the low to mid copy plasmid pSB3K3, and J23115.E0240, which has a weaker promotor than J23101.E0240. During the experiment, we could observe a typical growth curve for ''E.coli'' including lag, exponential, stationary and death phase. We could show that the fluorescence we measured is rather a function of each cell than the whole culture, since all cultures had comparable optical densities. | We introduced the three constructs in to ''E. coli'' cells and measured fluorescence as well as optical density of the liquid cultures over a period of 18 hours, using a spectrophotometer and a plate reader, respectively. The obtained results confirmed our hypothesis that the fluorescence of the BioBrick in the high copy plasmid pSB1C3, J23101.E0240, would exhibit a stronger signal than the constructs I20260, which is on the low to mid copy plasmid pSB3K3, and J23115.E0240, which has a weaker promotor than J23101.E0240. During the experiment, we could observe a typical growth curve for ''E.coli'' including lag, exponential, stationary and death phase. We could show that the fluorescence we measured is rather a function of each cell than the whole culture, since all cultures had comparable optical densities. | ||
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==Experimental Design== | ==Experimental Design== | ||
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Over a time span of 18 hours the optical density and fluorescence of cultures containing these BioBricks were measured every 2 hours using the spectrophotometer and plate reader, respectively. | Over a time span of 18 hours the optical density and fluorescence of cultures containing these BioBricks were measured every 2 hours using the spectrophotometer and plate reader, respectively. | ||
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==Expected Results== | ==Expected Results== | ||
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B0015 was used as our negative control as the insert only contains a terminator and no expression cassette for GFPmut3b, and therefore no fluorescence was expected. | B0015 was used as our negative control as the insert only contains a terminator and no expression cassette for GFPmut3b, and therefore no fluorescence was expected. | ||
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==Materials and Methods== | ==Materials and Methods== | ||
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As for the OD measurement, we used LB medium as our blank. Since samples for fluorescence measurement were acquired from the cuvettes for the OD measurement, sample processed in plate reader had the same dilutions. | As for the OD measurement, we used LB medium as our blank. Since samples for fluorescence measurement were acquired from the cuvettes for the OD measurement, sample processed in plate reader had the same dilutions. | ||
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==Results== | ==Results== | ||
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The development of fluorescence followed largely the pattern of the OD, but differed a lot in between the different cultures. J23101.E0240 exhibited fluorescence three times stronger than I20260, and about 10 times stronger than B0015 and J23115.E0240. The latter two did not differ in terms of fluorescent signal. | The development of fluorescence followed largely the pattern of the OD, but differed a lot in between the different cultures. J23101.E0240 exhibited fluorescence three times stronger than I20260, and about 10 times stronger than B0015 and J23115.E0240. The latter two did not differ in terms of fluorescent signal. | ||
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+ | {{Team:Aachen/BlockSeparator}} | ||
==Discussion== | ==Discussion== |
Revision as of 14:56, 3 October 2014
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