Team:Paris/August 21
From 2008.igem.org
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==Ligations from digestions from 20th== | ==Ligations from digestions from 20th== | ||
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[[Team:Paris/Notebook/Protocols#Transformation |Protocol]] | [[Team:Paris/Notebook/Protocols#Transformation |Protocol]] | ||
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Revision as of 14:11, 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; 13 clones were analysed by PCR PCR screeningreaction mixture (25 µL)
PCR screening programm
Electrophoresis
Results: *The clone of E0240 (S159.1) always have several bands amplified by PCR. Construction for synchronizationLigations
Promoter characterization plasmidsLigations from digestions from 20thTop 10 cells were used Protocol
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