Team:Paris/August 21
From 2008.igem.org
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='''Screening of the cloning of E0240 and FlhDC+promotor'''= | ='''Screening of the cloning of E0240 and FlhDC+promotor'''= | ||
- | We obtained | + | We obtained colonies isolated with the dilution 1/100. <br>''13 clones were analysed by PCR'' |
- | <br>13 clones were analysed by PCR | + | |
==PCR screening== | ==PCR screening== |
Revision as of 13:35, 6 September 2008
Cloning of FlhB promoterProtocol
Resuspension of 1 colony E.coli K12 strain MG 1655 in 100µl of water.
1µl of dNTP
Result
Construction for FIFOAim : Construction of pFlgA - YFP tripart (+/- LVA) "pFlgA-RBS-YFP-dbl ter" (pFlgA-E0430/E0432) DigestionDigestion
Gel Extraction
Measurement of the concentration of D166 & D167 purifiedProtocol (it's same that for Miniprep)
Ligation
Screening of the cloning of E0240 and FlhDC+promotorWe obtained colonies isolated with the dilution 1/100. PCR screeningreaction mixture (25 µL)
PCR screening programm
Electrophoresis
Construction for synchronizationLigations
Promoter characterization plasmidsLigations from digestions from 20thTop 10 cells were used
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