Team:Paris Saclay/Notebook/July/5

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='''Notebook : August 23'''=
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=='''Lab work'''==
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==='''A - Aerobic/Anaerobic regulation system'''===
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===='''Objective : obtaining Pfnr, NarK, NarG or NirB and RBS-LacZ-Term or RBS-AmilCP-Term in PSB3K3'''====
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(((((((((((((((===='''1 - Electrophoresis to check the digestion of Bba_J04450 by EcoRI/PstI'''====
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{|
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| style="width:350px;border:1px solid black;" |]]
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| style="width:350px;border:1px solid black;vertical-align:top;" |
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* Well 1 : 6µL DNA Ladder
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* Well 2 : 5µL of Bba_J04450 digested by EcoRI/PstI+1µl of 6X loading dye
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* Gel : 1%
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Expect sizes :
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* PSB3K3 : 2750 bp
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{|
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| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
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We can see anything. The digestion did work.
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|}))))))))))))))))))))))
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{{Team:Paris_Saclay/incl_fin}}

Revision as of 16:36, 22 September 2013

Navigation Home Team Project Labwork Notebook Human practices

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

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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Navigation Home Team Project Labwork Notebook Human practices

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====

]]
  • Well 1 : 6µL DNA Ladder
  • Well 2 : 5µL of Bba_J04450 digested by EcoRI/PstI+1µl of 6X loading dye
  • Gel : 1%

Expect sizes :

  • PSB3K3 : 2750 bp

We can see anything. The digestion did work.

))))))))))))))))))))))

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