Team:Paris Saclay/Notebook/July/5

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(Notebook : July 5)
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<u>Electrophoresis</u><br>
<u>Electrophoresis</u><br>
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*Well 1,2 : Not I
*Well 1,2 : Not I

Revision as of 09:04, 21 September 2013

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

PCRPS050813.jpg
  • Well 1,2 : Not I
  • Well 3,4 : Hpa I
  • Well 6,7 : Mlu I
  • gel 0.8%

It seemed that we had made some terrible mistakes. We continued and did another digestion the next week.




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