Team:Paris/August 26
From 2008.igem.org
(→Construction of pFlhB - mRFP Tripart (LVA+)) |
(→Miniprep and stock glycerol) |
||
Line 321: | Line 321: | ||
- | ='''Miniprep and stock | + | ='''Miniprep and stock glycerolof New Biobrick'''= |
- | |||
*[[Team:Paris/Notebook/Protocols#Glycerol stocks| Protocol stocks]] | *[[Team:Paris/Notebook/Protocols#Glycerol stocks| Protocol stocks]] | ||
{|border="1" style="text-align: center" | {|border="1" style="text-align: center" | ||
- | |''' | + | |'''Miniprep''' |
+ | |'''Strain''' | ||
|'''Biobricks''' | |'''Biobricks''' | ||
|'''Description''' | |'''Description''' | ||
|- | |- | ||
- | |S1 | + | |MP1.1 |
+ | |S1.1 | ||
|rowspan="2"| | |rowspan="2"| | ||
|rowspan="2"| [[Image:Icon_rbs.png]]<br>RBS | |rowspan="2"| [[Image:Icon_rbs.png]]<br>RBS | ||
|- | |- | ||
- | | | + | |MP1.2 |
+ | |S1.2 | ||
|- | |- | ||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | + | |MP1.1 | |
- | + | |S1.1 | |
- | + | ||
- | | | + | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
|rowspan="2"| | |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 | |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 | + | |MP1.2 |
+ | |S1.2 | ||
|- | |- | ||
- | |MP1 | + | |MP1.1 |
+ | |S1.1 | ||
|rowspan="2"| | |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 | |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 | + | |MP1.2 |
+ | |S1.2 | ||
|- | |- | ||
- | |MP1 | + | |MP1.1 |
+ | |S1.1 | ||
|rowspan="2"| | |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 | |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 | + | |MP1.2 |
+ | |S1.2 | ||
|- | |- | ||
- | |MP1 | + | |MP1.1 |
+ | |S1.1 | ||
|rowspan="2"| | |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 | |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 | + | |MP1.2 |
- | + | |S1.2 | |
- | + | ||
- | + | ||
- | | | + | |
- | + | ||
- | + | ||
|} | |} | ||
- | |||
- | |||
='''Construction of pFlhB - mRFP Tripart (LVA+)'''= | ='''Construction of pFlhB - mRFP Tripart (LVA+)'''= |
Revision as of 18:35, 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 glycerolof New BiobrickConstruction 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
|