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| - | == Results ==
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| - | [[Image:Montage.jpg|thumb|right|300px|Microscopy results of iGEMgfp upon 1% induction by subtilin]]
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| - | [[Image:Montage_pc.jpg|thumb|right|300px|Microscopy results of iGEMcherry upon 1% induction by subtilin]]
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| - | [[Image:Flow0%.jpg|thumb|right|300px|Flow cytometry results for 0% induction by subtilin.]]
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| - | [[Image:Flow1%.jpg|thumb|right|300px|Flow cytometry results for 1% induction by subtilin.]]
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| - | [[Image:Flow10%.jpg|thumb|right|300px|Flow cytometry results for 10% induction by subtilin.]]
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| - | To analyse the results of the wet lab transformations of the inserts into ''B. subtilis'', we used two methods: microscopy and flow cytometry.
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| - | ===Microscopy===
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| - | Microscopy work from 08.09.08 showed a difference in the level of flourescence of the iGEMgfp fluorescent cells (higher in 10% subtilin-induced cells compared to 0% subtilin-induced cells). However, there was little difference in the ''number'' of cells that fluoresced between the two cultures.
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| - | There was no difference in the number of fluorecent cells ''or'' the level of fluorescence between the 10% subtilin-induced and the 0% subtilin-induced iGEMcherry cells.
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| - | ===Flow cytometry===
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| - | Flow cytometry measures cell fluorescence and light scattering, cell-by-cell, allowing us to quantify our results and present them in graphical form. A sample of our cells engineered Bacillus subtilis cells were injected into the machine, which hydro-dynamically focusses the fluid. Lasers of a particular wavelength are directed onto the stream of fluid, and each particle which passes through the light beam will cause the laser to scatter in a different way. Cells which absorb and then re-emit the light, result in fluorescence (a higher energy state).
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| - | The detectors in the machine measure the scattering of light and any fluoresence which occurs, and the data is displayed as histograms and dot plots.
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| - | '''0% induction by subtilin''' (i.e in the absence of subtilin): mean fluoresence = 7.70
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| - | '''1% induction by subtilin:''' mean fluoresence = 14.77
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| - | '''10% induction by subtilin:''' the mean fluoresence = 21.95
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| - | These results show that the higher the concentraion of subtilin, the more GFP is expressed.
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| - | At 10% induction, the subtilin began to cause some of the cells to lyse- this can be seen by the two distinct populations of cells- one being the cells which have lysed, and the other being the remainder of intact cells. Since subtilin is an antibiotic, this was a good indication that the subtilin we had collected that day was of good quality.
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| - | </div>
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