Team:Paris Saclay/Notebook/August/6

From 2013.igem.org

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* Well 1 : 6µL DNA Ladder
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* Well 9 to 14 : 10µL BBa_K1155004+2µl of 6X loading dye
* Well 9 to 14 : 10µL BBa_K1155004+2µl of 6X loading dye
* Well 15 : 6µL DNA Ladder
* Well 15 : 6µL DNA Ladder
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* Well 1 : 6µL DNA Ladder
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* Well 2 to 7 : 10µL BBa_K1155004+2µl of 6X loading dye
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* Well 9 to 15 : 10µL BBa_K1155004+2µl of 6X loading dye
* Well 16 : 6µL DNA Ladder
* Well 16 : 6µL DNA Ladder
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* Well 17 to 22 : 10µL BBa_K1155004+2µl of 6X loading dye
 
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* Well 23 : 6µL DNA Ladder
 
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* Well 24 to 30 : 10µL BBa_K1155004+2µl of 6X loading dye
 
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* Gel : 1%
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Revision as of 23:25, 1 October 2013

Contents

Notebook : August 6

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155004, BBa_K1155005, BBa_K1155006

1 - Electrophoresis to check the Colony PCR products : BBa_K1155004, BBa_K1155005, BBa_K1155006

XiaoJing, Damir, Anaïs

  • BBa_K1155004 :
Psgel10608.jpgPsgel20608.jpg
  • Well 1 : 6µL DNA Ladder
  • Well 2 to 7 : 10µL BBa_K1155004+2µl of 6X loading dye
  • Well 8 : 6µL DNA Ladder
  • Well 9 to 14 : 10µL BBa_K1155004+2µl of 6X loading dye
  • Well 15 : 6µL DNA Ladder
  • Well 1 : 6µL DNA Ladder
  • Well 2 to 7 : 10µL BBa_K1155004+2µl of 6X loading dye
  • Well 8 : 6µL DNA Ladder
  • Well 9 to 15 : 10µL BBa_K1155004+2µl of 6X loading dye
  • Well 16 : 6µL DNA Ladder
  • Gel : 1%
  • BBa_K1155005 :
Psgel30608.jpgPsgel40608.jpg
  • Well 1 : 6µL DNA Ladder
  • Well 2 to 7 : 10µL BBa_K1155005+2µl of 6X loading dye
  • Well 8 : 6µL DNA Ladder
  • Well 9 to 14 : 10µL BBa_K1155005+2µl of 6X loading dye
  • Well 15 : 6µL DNA Ladder
  • Well 16 : 6µL DNA Ladder
  • Well 17 to 22 : 10µL BBa_K1155005+2µl of 6X loading dye
  • Well 23 : 6µL DNA Ladder
  • Well 24 to 30 : 10µL BBa_K1155005+2µl of 6X loading dye
  • Well 31 : 6µL DNA Ladder
  • Gel : 1%
  • BBa_K1155006 :
Psgel11208.jpgPsgel50608.jpg
  • Well 1 to 12 : 10µL BBa_K1155006+2µl of 6X loading dye
  • Well 13 : 6µL DNA Ladder
  • Well 14 to 26 : 10µL BBa_K1155006+2µl of 6X loading dye
  • Gel : 1%

Expected size :

  • NarK, NarG, NirB : 500 bp

We obtain fragments at the good size for all the colony. We will make a culture of DH5α with BBa_K1155004, BBa_K1155005 and BBa_K1155006. We will also sequence our plasmids.

2 - Liquid culture of DH5α with BBa_K1155004, BBa_K1155005 and BBa_K1155006

Xiaoing, Anaïs

Used quantities :

  • LB : 5mL
  • Chloramphénicol (1000X, 20µg/mL) : 5µL
  • BBa_K1155004, BBa_K1155005 and BBa_K1155006 : 25µL

We let the incubation over night at 37°C at 180 RPM.

We used colonies number 6, 7 and 8 for each promotor.

Objective : obtaining BBa_K1155007

1 - Electroelution of BBa_I732017 digested by EcoRI/SpeI

Nadia

Protocol : Electroelution

The electroelution was good. We will ligate RBS-LacZ with Term and PSB1C3.

Objective : obtaining biobricks in PSB3K3

1 - Extraction of BBa_J04450 from DH5α

Abdou

Protocol : Low copy plamid extraction

B - PCB sensing system

Objective : obtaining BBa_K1155002

1 - Sequence analysis for BBa_K1155002 in clones 4, 17 and 22

The sequence is good for each clone. We obtain a new biobrick : BBa_K1155002.


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