Team:Paris/August 26
From 2008.igem.org
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{{Paris/Calendar_Links|August 25|August 27}} | {{Paris/Calendar_Links|August 25|August 27}} | ||
+ | =Extraction of EnvZ* and OmpR* from ''E. coli'' genome= | ||
+ | ==Name of the PCR== | ||
+ | {|border="2" | ||
+ | |Name | ||
+ | |What's in ? | ||
+ | |Template DNA | ||
+ | |Oligo F | ||
+ | |Oligo R | ||
+ | |- | ||
+ | |PCR 147 | ||
+ | |EnvZ* | ||
+ | |S 120 | ||
+ | |O 126 | ||
+ | |O 127 | ||
+ | |- | ||
+ | |PCR 148 | ||
+ | |OmpR* | ||
+ | |S 119 | ||
+ | |O 138 | ||
+ | |O 139 | ||
+ | |- | ||
+ | |T 147 | ||
+ | |nothing | ||
+ | |no template | ||
+ | |O 126 | ||
+ | |O 127 | ||
+ | |- | ||
+ | |T 148 | ||
+ | |nothing | ||
+ | |no template | ||
+ | |O 138 | ||
+ | |O 139 | ||
+ | |} | ||
+ | ==Protocol== | ||
+ | |||
+ | '''Protocol''' | ||
+ | *10 µL Phusion HF Buffer 5X | ||
+ | *2.5 µL Oligo F 10 mM | ||
+ | *2.5 µL Oligo R 10 mM | ||
+ | *1 µL dNTP | ||
+ | *1 µL Template DNA | ||
+ | *33 µL H<sub>2</sub>O | ||
+ | |||
+ | '''PCR Programme''' | ||
+ | *98°C 30s Initial denaturation | ||
+ | *''CYCLE 30X'' | ||
+ | *98°C 10s Denaturation | ||
+ | *60°C 30s Annealing | ||
+ | *72°C 45s Elongation | ||
+ | *''END OF CYCLE'' | ||
+ | *72°C 5min Terminal Elongation | ||
+ | |||
+ | ==Resuts== | ||
+ | ===Electrophoresis settings=== | ||
+ | *Gel 1% agar | ||
+ | *Ladder 1kb | ||
=PCR= | =PCR= | ||
==PCR1== | ==PCR1== |
Revision as of 13:26, 26 August 2008
Extraction of EnvZ* and OmpR* from E. coli genomeName of the PCR
ProtocolProtocol
PCR Programme
ResutsElectrophoresis settings
PCRPCR1
Program: Gradient Vf=20µL PCR2
Program: promoter Vf=50µL PCR3
Program: PCRFliA1 Program: PCRFliA2 Program: PCRFliA3
Cloning of EnvZ*Concentration measurement by Biophotometer
Ligation
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