Team:Paris Saclay/Notebook/July/15

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Contents

Notebook : July 15

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155000

1 - Sequensis analysis of BBa_K1155000

Abdou

WE HAVE OUR FIRST BIOBRICK ! The sequence analysis was good. We call our biobrick BBa_K1155000.

Objective : obtaining BBa_K1155003, BBa_K1155007

1 - Liquid culture of BBa_B0015, BBa_B0017, BBa_I732019

Sheng, Zhou

Tranformations of 07/12/13 works. We will do a liquid culture of them.

We used 5 colonies of each plasmid.


B - PCB sensor system

Objective : obtaining BBa_K1155001, BBa_K1155002, BphR2

1 - Digestion of BBa_K1155001, BBa_K1155002 and BphR2 in pSB1C3 by EcoRI/PstI and SacII to check the size of fragments

Abdou

Used quantities :

  • EcoRI/PstI :
    • DNA : 8µL
    • Buffer Orange : 1.6µL
    • EcoRI : 1µL
    • PstI : 1µL
    • H2O : 4.4µL
  • SacII :
    • DNA : 8µL
    • Buffer Blue : 1.6µL
    • SacII : 1µL
    • H2O : 5.4µL

We let the digestion 1h30 at 37°C.

2 - PCR of digestion of BBa_K1155001, BBa_K1155002 and BphR2 in pSB1C3

Anaïs, Sheng, Zhou

Used quantities :

We mix our colonies in 20µL of H2O :

Used quantities :

  • BphA1 or BphR1 or BphR2 : 2µL
  • Mix : (it was divided in 2 tubes for each oligo combinaison with 23µL of mix in each tube)
    • VF or Pfnr_Up : 6µL
    • VR or Pfnr_Down or VR : 6µL
    • dNTP : 6µL
    • Buffer Dream Taq : 30µL
    • Dream Taq : 6µL
    • H2O : 246µL

PCR Program :

  • BphR1 :

PsPCRR11507.jpg

  • BphA1, BphR2 :

PsPCRR2A11507.jpg

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