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Team:Paris Saclay/Notebook/July/5
From 2013.igem.org
Contents |
Notebook : July 5
Summary:
For fnr promoter regulator system:
- Stocked 2 samples for each 2 colonies which contained AmilCP or BBa_K1155000.
- Performed restriction digest for amilCP and fnr from the medium seeded yesterday.
- Verification DNA quantity by NanoDrop.
- Performed electrophoresis and photo interpretation .
For PCBs sensor system:
- Stocked 2 samples of pseudomonas KE707.
- Received Primer BphA1P_up/down and BphR2_up/down.
Lab work
- A.aero/anaerobic regulation system
- 2.BioBrick RBS+LacZ+terminator in plasmid PSB1C3
- BioBrick RBS+amilCP+terminator in plasmid PSB1C3
Electrophoresis band size estimation
We used Clonemanager for band size estimation:
| Molecule | enzymes(Buffer) | Size |
| AmilCP clone | Nod I(Orange) | 2046+693bp |
| AmilCP clone | XHOI(Red) | 1842+892bp |
| BBa_K1155000 | XHOI(Red) | 1289+892bp |
| BBa_K1155000 | ECoR I(Orange) | 2151bp |
DNA quantification
Result(average):
- fnr promoter : 761ng/µl
- AmilCP : 229ng/µl
By using NanoDrop, we found some erreurs which could be due to device malfunctional issue.
Electrophoresis
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It seemed that we had made some terrible mistakes. We continued and did another digestion the next week.
| Previous day | Back to calendar | Next week |
Notebook : August 23
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Pfnr, NarK, NarG or NirB and RBS-LacZ-Term or RBS-AmilCP-Term in PSB3K3
(((((((((((((((====1 - Electrophoresis to check the digestion of Bba_J04450 by EcoRI/PstI====
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Expect sizes :
- PSB3K3 : 2750 bp
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We can see anything. The digestion did work. |
