Team:Paris Saclay/Notebook/September/28

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Notebook : August 28

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lab work

  • A - Aerobic/Anaerobic regulation system / B - PCB sensing system
    • 1 - Gel extraction of PSB1C3 Clean for Gibson digested Dnp1

Protocol : Gel extraction

    • 2 -Gel electrophoresis of product gel extraction.


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  • Well 1 : 6µL DNA Ladder
  • Well 2 : 3µL of product gel extraction+1µl of 6X loading dy


  • Gel : 1.0%
product quantification
We used the nano drop to measure the DNA in 260nm and we found its concentration
name PSB1C3 clean for Gibson
conc. 37.2ng/µl

i

  • We use it to do Gibson assembly .
    • 3- Gibson assembly.


  • RBS_BphR2_part1, BphR2_part2 and plasmid PSB1C3
  • FNR_part1, FNR part1 and plasmid PSB1C3
  • RBS_FNR part1, FNR_part2 and plasmid PSB1C3


Sample Volume:

  • Tube 1 : RBS_BphR2 part1(1ul), BphR2 part2(1ul) , plasmid PSB1C3(3ul),MIX Gibson (15 ul)
  • Tube 2 : FNR part1(1ul), FNR part2(1ul) , plasmid PSB1C3(3ul),MIX Gibson (15 ul)
  • Tube 3 : RBS_FNR part1(1ul), FNR part2(1ul) , plasmid PSB1C3(3ul),MIX Gibson (15 ul)

These 3 mixture is incubated at 50°C for up to one hour. Transformation and spread in 6 LB plates with Chlorenphenicol . Incubate at 37°C over night.

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