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Team:Paris Saclay/Notebook/August/12
From 2013.igem.org
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| - | *Well 1 : 6µL DNA Ladder | + | * Well 1 : 6µL DNA Ladder |
| - | *Well 2 : 5µL Bba_K1155000 digested by SpeI and PstI+1µl of 6X loading dye | + | * Well 2 : 5µL Bba_K1155000 digested by SpeI and PstI+1µl of 6X loading dye |
| - | *Well 3 : 5µL Bba_K1155007 digested by XBaI and PstI+1µl of 6X loading dye | + | * Well 3 : 5µL Bba_K1155007 digested by XBaI and PstI+1µl of 6X loading dye |
| - | *Well 4 : 5µL Bba_K1155003 digested by xBaI and PstI+1µl of 6X loading dye | + | * Well 4 : 5µL Bba_K1155003 digested by xBaI and PstI+1µl of 6X loading dye |
| - | *Gel : 0.8% | + | * Gel : 0.8% |
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Expected sizes : | Expected sizes : | ||
| - | *RBS_LacZ_Term : 3500 kb | + | * Pfnr : ... |
| - | *RBS_AmilCP_Term : ... | + | * RBS_LacZ_Term : 3500 kb |
| - | + | * RBS_AmilCP_Term : ... | |
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| - | *Well 1 : 6µL DNA Ladder | + | * Well 1 : 6µL DNA Ladder |
| - | *Well 2 : 5µL RBS-BphR2 part I+1µl of 6X loading dye | + | * Well 2 : 5µL RBS-BphR2 part I+1µl of 6X loading dye |
| - | *Well 3 : 5µL BphR2 part II+1µl of 6X loading dye | + | * Well 3 : 5µL BphR2 part II+1µl of 6X loading dye |
| - | *Well 4 : 5µL FNR part I+1µl of 6X loading dye | + | * Well 4 : 5µL FNR part I+1µl of 6X loading dye |
| - | *Well 5 : 5µL FNR part II+1µl of 6X loading dye | + | * Well 5 : 5µL FNR part II+1µl of 6X loading dye |
| - | *Well 6 : 5µL RBS-FNR part I+1µl of 6X loading dye | + | * Well 6 : 5µL RBS-FNR part I+1µl of 6X loading dye |
| - | *Well 4 : 5µL BphR2 part II+1µl of 6X loading dye | + | * Well 4 : 5µL BphR2 part II+1µl of 6X loading dye |
| - | *Gel : 0.8% | + | * Gel : 0.8% |
|} | |} | ||
Expected size | Expected size | ||
| - | *RBS-BphR2 part I : 197 kb | + | * RBS-BphR2 part I : 197 kb |
| - | *BphR2 part II : 790 kb | + | * BphR2 part II : 790 kb |
| - | *FNR part I : 597 kb | + | * FNR part I : 597 kb |
| - | *FNR part II : 200 kb | + | * FNR part II : 200 kb |
| - | *RBS-FNR part I : 615 kb | + | * RBS-FNR part I : 615 kb |
| - | *BphR2 part I : 178 kb | + | * BphR2 part I : 178 kb |
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Revision as of 10:39, 21 August 2013
Notebook : August 12
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining ...
1 - Digestion of Bba_K1155000 by SpeI and PstI, Bba_K1155007 and Bba_K1155003 by XBaI and PstI
Anaïs, Nadia, XiaoJing
- Bba_K1155000 :
- Buffer FD : 5µL
- H2O : 38µL
- DNA : 5µL
- SpeI : 1µL
- PstI : 1µL
- Bba_K1155007 :
- Buffer FD : 5µL
- H2O : 23µL
- DNA : 20µL
- XBal : 1µL
- PstI : 1µL
- Bba_K1155003 :
- Buffer FD : 5µL
- H2O : 33µL
- DNA : 10µL
- XBal : 1µL
- PstI : 1µL
2 - Electrophoresis to check the digestion of Bba_K1155000 by SpeI and PstI, Bba_K1155007 and Bba_K1155003 by XBalI and PstI
Nadia
| File:Psdigestion12.jpg|350px]] |
|
Expected sizes :
- Pfnr : ...
- RBS_LacZ_Term : 3500 kb
- RBS_AmilCP_Term : ...
|
We can't see any band for Bba_K1155000 digestion. The digestion failed. We will do it again. We obtain RBS-LacZ-Term and RBS-AmilCP-Term fragments. The digestion was good. We can now purify it. |
3 - Digestion of Bba_K1155000 by SpeI and PstI
Anaïs, Nadia
- Buffer FD : 5µL
- H2O : 38µL
- DNA : 5µL
- SpeI : 1µL
- PstI : 1µL
A - Aerobic/Anaerobic regulation system / B - PCB sensor system
Objective : obtaining FNR and BphR2 proteins
1 - Electrophoresis of PCR products : RBS-BphR2 part I, BphR2 part I, BphR2 part II, RBS-FNR part I, FNR part I and FNR part II
Damir
| [[]] |
|
Expected size
- RBS-BphR2 part I : 197 kb
- BphR2 part II : 790 kb
- FNR part I : 597 kb
- FNR part II : 200 kb
- RBS-FNR part I : 615 kb
- BphR2 part I : 178 kb
|
We can't see FNR Part I, FNR part II and BphR2 part i fragments at the good size. We will make the PCR again. We obtain RBS-BphR2 part I, BphR2 part II, RBS-FNR part I frangments at the right size thanks to the PCR. We will purify it. |
