Team:Paris Saclay/Notebook/July/5

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It seemed that we had made some terrible mistakes. We continued and did another digestion the next week.
*'''PCBs sensor system'''
*'''PCBs sensor system'''

Revision as of 15:54, 19 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. and performed a PCR
  • 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.

  • PCBs sensor system
    • 1.BioBrick BphR2(regulator) in plasmid PSB1C3




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