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Team:Paris Saclay/Notebook/July/4
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==='''A - Aerobic/Anaerobic regulation system'''=== | ==='''A - Aerobic/Anaerobic regulation system'''=== | ||
| - | ===='''Objective : obtaining | + | ===='''Objective : obtaining BBa_K1155000'''==== |
===='''1 - Estimation of Pndh* Colony PCR size fragments'''==== | ===='''1 - Estimation of Pndh* Colony PCR size fragments'''==== | ||
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| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" | | | style="border:1px solid black;padding:5px;background-color:#DEDEDE;" | | ||
| - | We obtain fragment at the right size in wells : 3, 4 , 8 , 9 12 and 13. We will digest our biobrick to confirm that Pndh* insert | + | We obtain fragment at the right size in wells : 3, 4 , 8 , 9 12 and 13. We will digest our biobrick to confirm that Pndh* insert pSB1C3 correctly. |
|} | |} | ||
| - | ===='''3 - Stock of | + | ===='''3 - Stock of BBa_K1155000'''==== |
Zhou | Zhou | ||
Used quantities : | Used quantities : | ||
| - | * | + | * BBa_K1155000 confirmed : 1 mL |
* Glycerol : 500µL glycerol. | * Glycerol : 500µL glycerol. | ||
We stocked them at -20°C. | We stocked them at -20°C. | ||
| - | ===='''4 - Extraction of | + | ===='''4 - Extraction of BBa_K1155000 from DH5α'''==== |
Anaïs, Sheng | Anaïs, Sheng | ||
| Line 57: | Line 57: | ||
Protocol : [[Team:Paris_Saclay/extraction|Hight copy plamid extraction]] | Protocol : [[Team:Paris_Saclay/extraction|Hight copy plamid extraction]] | ||
| - | ===='''5 - Digestion of | + | ===='''5 - Digestion of BBa_K1155000 by NotI, MluI, HpaI to check good insertion of Pndh* in pSB1C3'''==== |
Abdou | Abdou | ||
Used quantities : | Used quantities : | ||
| - | * | + | * BBa_K1155000 : 2µL |
* Bufer oranger : 2µL | * Bufer oranger : 2µL | ||
* NotI or MluI or HpaI : 0.5µL | * NotI or MluI or HpaI : 0.5µL | ||
| Line 69: | Line 69: | ||
We let the digestion 1h30 at 37°C. | We let the digestion 1h30 at 37°C. | ||
| - | ===='''6 - Culture of | + | ===='''6 - Culture of BBa_K1155000'''==== |
Anaïs, Sheng | Anaïs, Sheng | ||
| - | We made 2 cultures of bacterias transformed with | + | We made 2 cultures of bacterias transformed with BBa_K1155000 that show a fragments at the right size at electrophoresis. |
Latest revision as of 16:35, 4 October 2013
Notebook : July 4
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining BBa_K1155000
1 - Estimation of Pndh* Colony PCR size fragments
Abdou, Anaïs, Sheng
We used software gene manager to find the correct size of our fragments.
Estimated size :
- VF/VR primer -> Plasmid without Pndh* size : 277bp
- VF/Pfnr_down -> Plasmid with Pndh* size : 276bp
- Pfnr_Up/VR -> Plasmid with Pndh* size : 311bp
2 - Electrophoresis of Colony PCR products
Zhou
|
|
|
We obtain fragment at the right size in wells : 3, 4 , 8 , 9 12 and 13. We will digest our biobrick to confirm that Pndh* insert pSB1C3 correctly. |
3 - Stock of BBa_K1155000
Zhou
Used quantities :
- BBa_K1155000 confirmed : 1 mL
- Glycerol : 500µL glycerol.
We stocked them at -20°C.
4 - Extraction of BBa_K1155000 from DH5α
Anaïs, Sheng
Protocol : Hight copy plamid extraction
5 - Digestion of BBa_K1155000 by NotI, MluI, HpaI to check good insertion of Pndh* in pSB1C3
Abdou
Used quantities :
- BBa_K1155000 : 2µL
- Bufer oranger : 2µL
- NotI or MluI or HpaI : 0.5µL
- H2O : 15.5µL
We let the digestion 1h30 at 37°C.
6 - Culture of BBa_K1155000
Anaïs, Sheng
We made 2 cultures of bacterias transformed with BBa_K1155000 that show a fragments at the right size at electrophoresis.
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