Team:Paris/August 26
From 2008.igem.org
(→Ligation) |
(→Construction of pFlhB - mRFP Tripart (LVA+)) |
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*transformation of TOP10 competent cells with 5 µL of ligation product | *transformation of TOP10 competent cells with 5 µL of ligation product | ||
*spreading on LB plates + ampicilline and incubation overnight at 37°C | *spreading on LB plates + ampicilline and incubation overnight at 37°C | ||
+ | |||
+ | |||
+ | ='''Miniprep and stock glycerol'''= | ||
+ | |||
+ | ==Stock Glycerol of New Biobrick== | ||
+ | *[[Team:Paris/Notebook/Protocols#Glycerol stocks| Protocol stocks]] | ||
+ | |||
+ | {|border="1" style="text-align: center" | ||
+ | |'''Stock number''' | ||
+ | |'''Biobricks''' | ||
+ | |'''Description''' | ||
+ | |- | ||
+ | |S1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]]<br>RBS | ||
+ | |- | ||
+ | |S163.2 | ||
+ | |- | ||
+ | |S | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ ECFP+ LVA+ term | ||
+ | |- | ||
+ | |S1 | ||
+ | |- | ||
+ | |S1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ YFP+ LVA- term | ||
+ | |- | ||
+ | |S1 | ||
+ | |- | ||
+ | |S1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ YFP LVA+ term | ||
+ | |- | ||
+ | |S1 | ||
+ | |- | ||
+ | |S1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ ECFP LVA- term | ||
+ | |- | ||
+ | |S1 | ||
+ | |- | ||
+ | |S1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ mRFP LVA+ term | ||
+ | |- | ||
+ | |S1 | ||
+ | |} | ||
+ | |||
+ | ==Miniprep of New Biobricks== | ||
+ | *[[Team:Paris/Notebook/Protocols#Minipreps (Kit_Qiagen)| Protocol Miniprep]] | ||
+ | {|border="1" style="text-align: center" | ||
+ | |'''Miniprep''' | ||
+ | |'''Biobricks''' | ||
+ | |'''Description''' | ||
+ | |- | ||
+ | |MP1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]]<br>RBS | ||
+ | |- | ||
+ | |MP1 | ||
+ | |- | ||
+ | |MP1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ ECFP+ LVA+ term | ||
+ | |- | ||
+ | |MP1 | ||
+ | |- | ||
+ | |MP1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ YFP+ LVA- term | ||
+ | |- | ||
+ | |MP1 | ||
+ | |- | ||
+ | |MP1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ YFP LVA+ term | ||
+ | |- | ||
+ | |MP1 | ||
+ | |- | ||
+ | |MP1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ ECFP LVA- term | ||
+ | |- | ||
+ | |MP1 | ||
+ | |- | ||
+ | |MP1 | ||
+ | |rowspan="2"| | ||
+ | |rowspan="2"| [[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]<br>RBS+ mRFP LVA+ term | ||
+ | |- | ||
+ | |MP1 | ||
+ | |} | ||
+ | |||
Revision as of 18:05, 26 August 2008
Extraction of EnvZ* and OmpR* from E. coli genomeName of the PCR
ProtocolProtocol
PCR Programme
ResutsElectrophoresis settings
Results of the electrophoresis
Conclusion : All the PCR worked perfectly well ! Cleaning of the PCR productsThe cleaned PCR products are stored in the freezer overnight.
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
Miniprep and stock glycerolStock Glycerol of New Biobrick
Miniprep of New Biobricks
Construction of pFlhB - mRFP Tripart (LVA+)Aim : Construction of "pFlhB-RBS-mRFP-dbl ter" (pFlhB-I732078) We can only do the construction with mRFP Tripart (LVA+) because the stable strain with the Biobricks I732011 (mRFP Tripart LVA-) don't to growth. DigestionDigestion
Gel Verification
|