"
Team:Paris Saclay/Notebook/August/1
From 2013.igem.org
(Difference between revisions)
| Line 30: | Line 30: | ||
===='''2 - Gibson assembly.'''==== | ===='''2 - Gibson assembly.'''==== | ||
| - | Xiaojing | + | Abdou, Xiaojing |
Used quantities : | Used quantities : | ||
| Line 74: | Line 74: | ||
[[File:PsPCRBSBphR23007.jpg|400px]] | [[File:PsPCRBSBphR23007.jpg|400px]] | ||
| + | |||
| + | ===='''3 - Electrophoresis of the PCR products : RBS-BphR2 Part I'''==== | ||
| + | |||
| + | Xavier, XiaoJing | ||
| + | |||
| + | {| | ||
| + | | style="width:350px;border:1px solid black;" |File:Ps.jpg|350px]] | ||
| + | | style="width:350px;border:1px solid black;vertical-align:top;" | | ||
| + | * Well 1 : 6µL of DNA Ladder | ||
| + | * Well 2 : 5µL of RBS-BphR2 Part I+1µl of 6X loading dye | ||
| + | * Gel : 0.8% | ||
| + | |} | ||
| + | |||
| + | Expected sizes : | ||
| + | * RBS-BphR2 Part I : | ||
| + | |||
| + | {| | ||
| + | | style="border:1px solid black;padding:5px;background-color:#DEDEDE;" | | ||
| + | CONCLUSION !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! | ||
| + | |} | ||
===='''4 - Gel purification of PCR of : RBS-BphR2 Part I'''==== | ===='''4 - Gel purification of PCR of : RBS-BphR2 Part I'''==== | ||
Revision as of 21:31, 15 September 2013
Notebook : August 1
Lab work
A - Aerobic/Anaerobic regulation system / B - PCB sensing system
Objective : obtaining FNR and BphR2 proteins
1 - Gel purification of PCR products : BphR2 Part I, BphR2, Part II, FNR Part I, FNR Part II, RBS-FNR Part I, PSB1C3
Xavier
Protocol : Gel purification
Nanodrop :
- BphR2 Part I : 44 ng/µL
- BphR2 Part II : 64 ng/µL
- FNR Part I : 147 ng/µL
- FNR Part II : 140 ng/µL
- RBS-FNR Part I : 167 ng/µL
- PSB1C3 : 159 ng/µL
|
CONCLUSION !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! |
2 - Gibson assembly.
Abdou, Xiaojing
Used quantities :
- BphR2 :
- PSB1C3 : 2µL
- BphR2 Part I : 1µL
- BphR2 Part II : 1µL
- Gibson mix : 15µL
- H2O : 1µL
- FNR :
- PSB1C3 : 2µL
- FNR Part I : 1µL
- FNR Part II : 1µL
- Gisbon mix : 15µL ???????????????
- H2O : 1µL
- RBS-FNR :
- PSB1C3 : 2µL
- RBS-FNR Part I : 1µL
- FNR Part II : 1µL
- Gibson mix : 15µL ????????????
- H2O : 1µL
We let these mix at 50°C during 1h.
3 - PCR of : RBS-BphR2 Part I
Abdou
Used quantities :
- Oligo 54F : 1µL
- Oligo 55R : 1µL
- Buffer phusion : 5µL
- DNA : 0.25µL
- dNTP : 1µL
- Phusion : ... µL 5 ?????????
- H2O : 36.5µL
PCR Program :
3 - Electrophoresis of the PCR products : RBS-BphR2 Part I
Xavier, XiaoJing
| File:Ps.jpg|350px]] |
|
Expected sizes :
- RBS-BphR2 Part I :
|
CONCLUSION !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! |
4 - Gel purification of PCR of : RBS-BphR2 Part I
????????
