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Team:Newcastle University/Conclusions
From 2008.igem.org
Newcastle University
GOLD MEDAL WINNER 2008
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Results
To analyse the results of the wet lab transformations of the inserts into B. subtilis, we used two methods: microscopy and flow cytometry.
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.
There was no difference in the number of fluorecent cells or the level of flourescence between the 10% subtilin-induced and the 0% subtilin-induced iGEMcherry cells.
Flow cytometry
Flow cytometry allows us to quantify our results and present them in graphical form. A sample of cells our engineered Bacillus subtilis cells were injected into the machine which hydro-dynamically focusses the fluid. Lasers are directed onto the stream of fluid, and each particle which passes through the light beam will cause the laser to scatter in a particular way. Fluorescent chemicals are excited to a higher energy state.
The detectors in the machine measure the scattering of light and any flourescence which occurs.
0% induction by subtilin (i.e in the absence of subtilin): mean flourescence = 7.70
1% induction by subtilin: mean flourescence = 14.77
10% induction by subtilin: the mean flourescence = 21.95
These results show that the higher the concentraion of subtilin, the more GFP is expressed.
