Team:Paris Saclay/Notebook/July/5

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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.


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.







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Notebook : August 23

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining Bba_K1155000

1 - Electrophoresis of the digestion of Bba_K1155000 by NotI, HpaI, MluI

Sheng, Zhou

PCRPS050813.jpg
  • Well 1, 2 : 5µL of Bba_K1155000 digested by Not I+1µL of 6X loading dye
  • Well 3, 4 : 5µL of Bba_K1155000 digested by Hpa I+1µL of 6X loading dye
  • Well 5 : 6µL of DNA Ladder
  • Well 6, 7 : 5µL of Bba_K1155000 digested by Mlu I+1µL of 6X loading dye
  • Gel 0.8%

We didn't obtain fragments at the right size. We will do digestions again.

2 - Extraction of Bba_K1155000 and Bba_K592009 from DH5α

Abdou

Protocol : Hight copy plamid extraction

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