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Team:Paris Saclay/Notebook/August/6
From 2013.igem.org
(Difference between revisions)
(→1 - Electrophoresis to check the Colony PCR products : BBa_K1155004, BBa_K1155005, BBa_K1155006) |
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* Well 1 : 6µL DNA Ladder | * Well 1 : 6µL DNA Ladder | ||
| - | * Well 2 to 7 : 10µL BBa_K1155004+2µl of 6X loading dye | + | * Well 2 to 7 : 10µL BBa_K1155004 + 2µl of 6X loading dye |
* Well 8 : 6µL DNA Ladder | * Well 8 : 6µL DNA Ladder | ||
| - | * Well 9 to 15 : 10µL BBa_K1155004+2µl of 6X loading dye | + | * Well 9 to 15 : 10µL BBa_K1155004 + 2µl of 6X loading dye |
* Well 16 : 6µL DNA Ladder | * Well 16 : 6µL DNA Ladder | ||
* Well 1 : 6µL DNA Ladder | * Well 1 : 6µL DNA Ladder | ||
| - | * Well 2 to 7 : 10µL BBa_K1155004+2µl of 6X loading dye | + | * Well 2 to 7 : 10µL BBa_K1155004 + 2µl of 6X loading dye |
* Well 8 : 6µL DNA Ladder | * Well 8 : 6µL DNA Ladder | ||
| - | * Well 9 to 14 : 10µL BBa_K1155004+2µl of 6X loading dye | + | * Well 9 to 14 : 10µL BBa_K1155004 + 2µl of 6X loading dye |
* Well 15 : 6µL DNA Ladder | * Well 15 : 6µL DNA Ladder | ||
* Gel : 1% | * Gel : 1% | ||
Revision as of 18:29, 3 October 2013
Notebook : August 6
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining BBa_K1155004, BBa_K1155005, BBa_K1155006
1 - Electrophoresis to check the PCR Colonies products : BBa_K1155004, BBa_K1155005, BBa_K1155006
XiaoJing, Damir, Anaïs
- BBa_K1155004 :
 
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- BBa_K1155005 :
 
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- BBa_K1155006 :
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Expected size :
- NarK, NarG, NirB : 500 bp
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We obtain fragments at the good size for all the colony. We will make a culture of DH5α with BBa_K1155004, BBa_K1155005 and BBa_K1155006. We will also sequence our plasmids. |
2 - Liquid culture of DH5α with BBa_K1155004, BBa_K1155005 and BBa_K1155006
Xiaoing, Anaïs
Used quantities :
- LB : 5mL
- Chloramphénicol (1000X, 20µg/mL) : 5µL
- BBa_K1155004, BBa_K1155005 and BBa_K1155006 : 25µL
We let the incubation over night at 37°C at 180 RPM.
We used colonies number 6, 7 and 8 for each promotor.
Objective : obtaining BBa_K1155007
1 - Electroelution of BBa_I732017 digested by EcoRI/SpeI
Nadia
Protocol : Electroelution
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The electroelution was good. We will ligate RBS-LacZ with Term and pSB1C3. |
Objective : obtaining biobricks in pSB3K3
1 - Extraction of BBa_J04450 from DH5α
Abdou
Protocol : Low copy plamid extraction
B - PCB sensing system
Objective : obtaining BBa_K1155002
1 - Sequence analysis for BBa_K1155002 in clones 4, 17 and 22
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The sequence is good for each clone. We obtain a new biobrick : BBa_K1155002. |
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