"
Team:Paris Saclay/Notebook/August/19
From 2013.igem.org
(Difference between revisions)
| Line 9: | Line 9: | ||
===='''Objective : to obtain ... '''==== | ===='''Objective : to obtain ... '''==== | ||
| - | + | ====1 - Gel purification of the digestion of Bba_K1155003 and Bba_K1155007 by XBaI/PstI==== | |
Nadia | Nadia | ||
| Line 23: | Line 23: | ||
|} | |} | ||
| - | + | ====2 - EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term==== | |
Nadia, XiaoJing | Nadia, XiaoJing | ||
| Line 39: | Line 39: | ||
|} | |} | ||
| - | + | ====3 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3==== | |
XiaoJing | XiaoJing | ||
| Line 97: | Line 97: | ||
===='''Objective : obtaining FNR and BphR2 proteins (Gibson assembly) '''==== | ===='''Objective : obtaining FNR and BphR2 proteins (Gibson assembly) '''==== | ||
| - | + | ====1 - Electrophoresis gel of PCR products : BphR2 Part I==== | |
Damir | Damir | ||
| Line 117: | Line 117: | ||
|} | |} | ||
| - | + | ====2 - Gel purification of PCR products : BphR2 Part I ==== | |
Damir | Damir | ||
Revision as of 19:41, 11 September 2013
Contents |
Notebook : August 19
Lab work
A - Aerobic/Anaerobic regulation system
Objective : to obtain ...
1 - Gel purification of the digestion of Bba_K1155003 and Bba_K1155007 by XBaI/PstI
Nadia
Protocol : Gel purification
Nanodrop :
- RBS-LacZ-Term : 59.6ng/µL
- RBS-AmilCP-Term : 21.3ng/µL
|
Concentrations are very low so we try to precipitate the DNA again. |
2 - EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term
Nadia, XiaoJing
Protocol : EtOH precipitation Protocole ou à rédiger ici ?????????
We used 20µL of DNA.
Nanodrop :
- RBS-LacZ-Term : 38.4ng/µL
- RBS-AmilCP-Term : 28.7ng/µL
|
CONCLUSION |
3 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3
XiaoJing
Used quantities :
- NirB and RBS-LacZ-Term :
- NirB : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NirB and RBS-AmilCP-Term :
- NirB : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarG and RBS-LacZ-Term :
- NarG : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarG and RBS-AmilCP-Term :
- NarG : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarK and RBS-LacZ-Term :
- Nar K : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarK and RBS-AmilCP-Term :
- NarK : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
A - Aerobic/Anaerobic regulation system / B - PCB sensor system
Objective : obtaining FNR and BphR2 proteins (Gibson assembly)
1 - Electrophoresis gel of PCR products : BphR2 Part I
Damir
| [[]] |
|
Expected size :
- BphR2 Part I : 178 kb
|
We obtain a frangment at the right size. We can purify it. |
2 - Gel purification of PCR products : BphR2 Part I
Damir
Protocol : Gel purification
Nanodrop :
- BphR2 : 57.2ng/µL
|
CONCLUSION |
Meeting with Evry
