Team:Paris Saclay/Notebook/August/6

From 2013.igem.org

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==='''A - Aerobic/Anaerobic regulation system / B - PCB sensing system'''===
==='''A - Aerobic/Anaerobic regulation system / B - PCB sensing system'''===
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====1 - PCR product( made the 08/01/2013)digestion to degrade the backbone psB1C3
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{{Team:Paris_Saclay/incl_fin}}
{{Team:Paris_Saclay/incl_fin}}

Revision as of 13:02, 11 August 2013

Contents

Notebook : August 6

Lab work

A - Aerobic/Anaerobic regulation system

Obtaining activator FNR promoter (narK, nirB, narG)

1 - Gel electrophoresis of the colony PCR products ( made the 08/05/2013)on 25 colonies

XiaoJing, Damir, Anaïs

  • NarK promoter in psB1C3
IMAGE
  • Wells 1 to 25 : 10µl sample
  • Well between 12 and 13 : 6µl gene ruler
  • Gel : 1.5%

Expected size : 500 bp

We obtained fragments of the right size. Our backbone contains the Biobrick.
  • NirB promoter in psB1C3
IMAGE
  • Before well 1  : 6µl gene ruler
  • Wells 1 to 6 : 10µl sample
  • Between 6 and 7: 6µl gene ruler
  • Wells 7 to 12 : 10µl sample
  • After well 12 :6µl gene ruler
  • Before well 13  : 6µl gene ruler
  • Wells 13 to 18 : 10µl sample
  • Between 18 and 19: 6µl gene ruler
  • Wells 19 to 25 : 10µl sample
  • After well 25 :6µl gene ruler
  • Gel : 1.5%

Expected size : 500 bp

We obtained fragments of the right size. Our backbone contains the Biobrick.
  • NarG promoter in psB1C3
IMAGE
  • Before well 1  : 6µl gene ruler
  • Wells 1 to 6 : 10µl sample
  • Between 6 and 7: 6µl gene ruler
  • Wells 7 to 12 : 10µl sample
  • After well 12 :6µl gene ruler
  • Before well 13  : 6µl gene ruler
  • Wells 13 to 18 : 10µl sample
  • Between 18 and 19: 6µl gene ruler
  • Wells 19 to 25 : 10µl sample
  • After well 25 :6µl gene ruler
  • Gel : 1.5%

Expected size : 500 bp

We obtained fragments of the right size. Our backbone contains the Biobrick.

2 - Samples culture

Xiaoing, Anaïs

We put in culture the 6, 7, 8 samples for every promoter in 5 ml LB + 5 µl Chlorenphenicol (1000x,20µg/ml) Incubator over night to 310,15K at 180 RPM

B - PCB sensing system

Obtaining the BphA1 promoter

====1 - Sequence analysis for BB pBphA1 in psB1C3 5 ( clone 4, 17 and 22 )

IMAGE

A - Aerobic/Anaerobic regulation system / B - PCB sensing system

====1 - PCR product( made the 08/01/2013)digestion to degrade the backbone psB1C3