IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/18: Difference between revisions
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Revision as of 04:38, 5 September 2008
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Check promotersOn Friday, PCR out promoters, we want to check them. - Run a gel - Results : good size of bands!!! Test Ery efficiencySince all Ery plates grew, we want to check the antibiotic stock. - New Ery plate of IA751 (they should die) - New LB stock (Ery5) with IA751 Single colony plates- New plates (Cm5 + Amp100) with single colony of ECE188 (from 14/08), ECE189 (from 13/08) and ECE190 (from 14/08) - Grow the same single colony into LB with antibiotic Transformation of glycerol stocks of B.S.The result from the last transformation of glycerol stocks was negative> We tried again with two different protocols.
- Spin the glycerol stock, throw out the supernatant (just keep about 100μL), add 0.5μL of Medium B - Incubate about 1h15 - Add 0.6μg of ECE166 (because we managed to transform BS with this vector) - Incubate 1h30 - Plate on Cm5 plates (DNA less control and transformed cells)
- Spin the glycerol stock, throw out the supernatant (just keep about 100μL), add 0.5μL of Medium B - Add 0.6μg of ECE166 (because we managed to transform BS with this vector) - Incubate 30min - Plate on Cm5 plates (transformed cells)
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Transformation of AgrA and AgrC
AgrA and C gell-extracted ligation was transformed into Top 10 cells using standard protocol. Puc9 was used as a positive control.