Team:Paris Saclay/Notebook/July/5

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(Difference between revisions)
(Lab work)
(Notebook : July 5)
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*Performed electrophoresis and photo interpretation .
*Performed electrophoresis and photo interpretation .
For PCBs sensor system:
For PCBs sensor system:
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*Stocked 2 samples of pseudomonas KE707. and performed a PCR
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*Stocked 2 samples of pseudomonas KE707.
*Received Primer BphA1P_up/down and BphR2_up/down.
*Received Primer BphA1P_up/down and BphR2_up/down.
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It seemed that we had made some terrible mistakes. We continued and did another digestion the next week.
It seemed that we had made some terrible mistakes. We continued and did another digestion the next week.
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*'''PCBs sensor system'''
 
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**''1.BioBrick BphR2(regulator) in plasmid PSB1C3''
 

Revision as of 15:55, 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.
  • 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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