"
Team:Paris Saclay/Notebook/August/22
From 2013.igem.org
Notebook : August 22
Lab work
A - Aerobic/Anaerobic regulation system
Objective : characterize BBa_K1155000, BBa_K1155004, BBa_K1155005, BBa_K1155006
1 - Plasmid extraction of BBa_K1155000 from DH5α
Nguyen
Protocol : High-copy plamid extraction
Nanodrop :
- BBa_K1155000 : 175ng/µL
|
The extraction was good. We will digested the plasmid. |
2 - Digestion of BBa_K1155000 by SpeI
Used quantities :
- BBa_K1155000 : 10µL
- Buffer FD : 2µL
- SpeI : 2µL
- H2O : 6 µL
We let digestions at 37°C during 10 minutes.
Objective : obtaining Pfnr, NarK, NarG or NirB and RBS-LacZ-Term or RBS-AmilCP-Term in pSB3K3
1 - Gel purification of the digestion of BBa_J04450 by EcoRI/PstI
XiaoJing
Protocol : Gel purification
Nanodrop :
- pSB3K3 : 4ng/µL
|
The Nanodrop gives us a very few quantity of pSB3K3 so we decided to check it with a first electrophoresis. |
2 - Electrophoresis of gel purification of the digestion of BBa_J04450 by EcoRI/PstI
XiaoJing
|
|
Expect sizes :
- pSB3K3 : 2750 bp
|
We can't see anything with the fisrt electrophoresis that's why we made an EtOH precipitation. |
3 - Ethanol precipitation of the digestion of BBa_J04450 by EcoRI/PstI
Protocol : EtOH precipitation
We used 34µL of DNA.
A - Aerobic/Anaerobic regulation system / B - PCB sensor system
Objective : obtaining BphR2 protein
1 - PCR of BphR2 Part I
Damir
Used quantities :
- Oligo 54F : 2µL
- Oligo 55R : 2µL
- DNA : 1µL
- Buffer Phusion : 10µL
- dNTP : 1µL
- Phusion : 1µL
- DMS0 : 2µL
- H2O : 31µL
PCR program :
2 - Electrophoresis of PCR product : BphR2 Part I
Damir
|
|
Expected sizes :
- BphR2 Part I : 178 kb
|
We obtained fragments at the right size. We can purify it. |
3 - Gel purification of PCR product : BphR2 Part I
Damir
Protocol : Gel purification
Nanodrop :
- RBS-BphR2 Part I, tube 1 : 42ng/µL
- RBS-BphR2 Part I, tube 2 : 75ng/µL
| Previous day | Back to calendar | Next day |
