"
Team:Paris Saclay/Notebook/August/5
From 2013.igem.org
(Difference between revisions)
| Line 9: | Line 9: | ||
===='''Objective : obtaining Bba_K1155003'''==== | ===='''Objective : obtaining Bba_K1155003'''==== | ||
| - | ===='''1 - Digestion of Bba_K1155003 by EcoRI/PstI'''==== | + | ===='''1 - Digestion of Bba_K1155003 by EcoRI/PstI to check the ligation between RBS-Amil CP,Term and PSB1C3 '''==== |
Nadia, XiaoJing | Nadia, XiaoJing | ||
| Line 20: | Line 20: | ||
* H2O : 20µL | * H2O : 20µL | ||
| - | We let the digestion at 37°C during | + | We let the digestion at 37°C during 15 minutes. |
===='''2 - Electrophoresis of the digestion of Bba_K1155003'''==== | ===='''2 - Electrophoresis of the digestion of Bba_K1155003'''==== | ||
| Line 39: | Line 39: | ||
* Gel : 1% | * Gel : 1% | ||
|} | |} | ||
| + | |||
| + | Expected sizes : | ||
| + | * RBS_Amil CP-Term : | ||
| + | * PSB1C3 : | ||
===='''Objective : obtaining Bba_K1155007'''==== | ===='''Objective : obtaining Bba_K1155007'''==== | ||
| Line 44: | Line 48: | ||
===='''1 - Digestion of Bba_I732017 by EcoRI/SpeI'''==== | ===='''1 - Digestion of Bba_I732017 by EcoRI/SpeI'''==== | ||
| - | Damir, Nadia | + | Abdou, Damir, Nadia, |
Used quantities : | Used quantities : | ||
| Line 61: | Line 65: | ||
| style="width:350px;border:1px solid black;" |[[]] | | style="width:350px;border:1px solid black;" |[[]] | ||
| style="width:350px;border:1px solid black;vertical-align:top;" | | | style="width:350px;border:1px solid black;vertical-align:top;" | | ||
| - | * Well 1 : | + | * Well 1 : 40µL of Bba_I732017 digested by EcoRI/SpeI+8µL 6X loading dye |
| - | + | * Gel : 1.5% | |
| - | * Gel : | + | |
|} | |} | ||
| Line 72: | Line 75: | ||
{| | {| | ||
| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" | | | style="border:1px solid black;padding:5px;background-color:#DEDEDE;" | | ||
| - | We | + | We obtain fragments at the right size. We will make an elector-elution to extract RBS-LacZ. |
|} | |} | ||
Revision as of 17:27, 20 September 2013
Notebook : August 5
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155003
1 - Digestion of Bba_K1155003 by EcoRI/PstI to check the ligation between RBS-Amil CP,Term and PSB1C3
Nadia, XiaoJing
Used quantities :
- Bba_K1155003 : 5µL
- EcoRI FD : 1µL
- PstI FD : 1µL
- Buffer FD : 3µL
- H2O : 20µL
We let the digestion at 37°C during 15 minutes.
2 - Electrophoresis of the digestion of Bba_K1155003
Damir
| File:Ps.jpg|350px]] |
|
Expected sizes :
- RBS_Amil CP-Term :
- PSB1C3 :
Objective : obtaining Bba_K1155007
1 - Digestion of Bba_I732017 by EcoRI/SpeI
Abdou, Damir, Nadia,
Used quantities :
- Bba_I732017 : 41µL
- Buffer FD : 5µL
- EcoRI : 2µL
- SpeI : 2µL
We let the digestion at 1h30 at 37°C.
2 -Electrophoresis to check the digestion of Bba_I732017 by EcoRI/SpeI
Damir, Nadia
| [[]] |
|
Expected sizes :
- RBS-LacZ : 3093 bp
- PSB1A2 : 2079 bp
|
We obtain fragments at the right size. We will make an elector-elution to extract RBS-LacZ. |
Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006
1 - Colony PCR of Bba_K1155004, Bba_K115005, Bba_K1155006 in DH5α
Damir, Nadia, XiaoJing
COLONIES PIQUEES DANS 10µL d'eau par Tube !!!!!!!!!!!!!
Used quantities :
- DNA : 2µL
- Mix : (it was divided in 25 tubes for each promotor with 23µL of mix in each on)
- Oligo 44 : 3.5µL
- Oligo 45 : 3.5µL
- Buffer Dream Taq : 70µL
- dNTP : 28µL
- Dream Taq : 5µL
- H2O : 590µL
PCR Program :
