Team:Paris/August 21
From 2008.igem.org
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==Protocol== | ==Protocol== | ||
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* '''Preparation of the template''' : | * '''Preparation of the template''' : | ||
Resuspension of 1 colony ''E.coli'' K12 strain MG 1655 in 100µl of water. | Resuspension of 1 colony ''E.coli'' K12 strain MG 1655 in 100µl of water. | ||
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* '''Preparation of PCR mix''' : | * '''Preparation of PCR mix''' : | ||
- | + | 1µl of dNTP | |
<br> 10µl Buffer Phusion 5X | <br> 10µl Buffer Phusion 5X | ||
<br> 2.5µl O 108 | <br> 2.5µl O 108 | ||
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<br> 1µl Phusion | <br> 1µl Phusion | ||
<br> 33µL pure water | <br> 33µL pure water | ||
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==Result== | ==Result== |
Revision as of 20:08, 21 August 2008
Cloning of FlhB promoterProtocol
Resuspension of 1 colony E.coli K12 strain MG 1655 in 100µl of water.
1µl of dNTP
Result[[Image:KR00019b.jpg| thumb| Vérification of pFlhB]
Construction of pFlgA - YFP tripart (+/- LVA)Aim : Construction of "pFlgA-RBS-YFP-dbl ter" (pFlgA-E0430/E0432) DigestionGel ExtractionProtocol [[Image:KR00019b.jpg| thumb|Gel Extraction of D166-D167]
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 single colonies with the dilution 100.
PCR screeningreaction mixture (25 µL)
PCR screening programm
Electrophoresis
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