Biomod/2011/TUM/TNT/LabbookA/2011/09/06: Difference between revisions
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<h2>Size exclusion filter purification of BM_2_15_65 and BM_2_5D_H3_ML </h2> | <h2>Size exclusion filter purification of BM_2_15_65 and BM_2_5D_H3_ML </h2> | ||
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6th Sep 2011
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Main project page
Size exclusion filter purification of BM_2_15_65 and BM_2_5D_H3_MLFolded samples BM_2_15_65 (2011/08/23) and BM_2_5D_H3_ML (2011/08/23) filtered in Amicon filters (100 kDa) with 0.5 x TBE + 11 mM MgCl2 Sample filtering
Control of filtration outcomeFigure XX shows a 50 ml 2% Agarose-Gel with EtBr Figure XX: From left to right: 1kb ladder, p7560, filtered BM_2, BM_2_15_65, BM_2_5D_H3_ML, 1st flowthrough, 2nd flowthrough
Size exclusion filter purification of folded BM_1_15_65Folded sample BM_1_1565(2011/08/23) in Amicon (100 kDa) with 1 x FOBxM20 Sample filtering
Control of filtration outcomeFigure XX shows a 125 ml 2% Agarose-Gel with EtBr Figure XX: 1kb ladder, p7560, BM_1_15_65 5x filtered, BM_1_1565 unfiltered, 1. flowthrough, 2. flowthrough, 3. flowthrough, 4. flowthrough, 5. flowthrough, ...Martys stuff ConclusionAfter 5 times filtering, no more staples can be seen in the agarose gel, the samples can be considered pure enough for further analysis. Some origamis are lost due to the filtering, but this loss still seems to be smaller than that when purifying with preparative agarose gel electrophoresis. Photometric spectroscopy of MH_255_647 and MH_256_550 with EtBrThis experiment is done to compare the results from the real time PCR experiments and photometric spectroscopy. Sample preparationPreliminary considerations
stocks
Labeled DNA
Control: Unlabled DNA
Hybridisation:
ResultsThe experiments done with the spectrometer failed. No signal was detectable at all. The concentrations of the samples were below the detection limit. Since the concentration of the origamis will be even lower this method is not usable for bulk measurements. |