Team:Paris Saclay/Notebook/August/6

From 2013.igem.org

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We obtain fragments at the good size for all the colony. ENSUITE ?????????
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We obtain fragments at the good size for all the colony. ?????????
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====2 - Samples culture====
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====2 - Culture of DH5α with Bba_K1155004, Bba_K1155005 and Bba_K1155006 ====
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Xiaoing, Anaïs
Xiaoing, Anaïs
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We put in culture the 6, 7, 8 samples for every promoter in 5 ml LB + 5 µl Chlorenphenicol (1000x,20µg/ml)
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Used quantities :
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Incubator over night to 310,15K at 180 RPM
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* LB : 5mL
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* Chloramphénicol (1000X, 20µg/mL) : 5µL
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* Bba_K1155004, Bba_K1155005 and Bba_K1155006 : 25µL ?????????
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==='''B - PCB sensing system'''===
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We let the incubation over night at 37°C at 180 RPM.
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===='''Obtaining the BphA1 promoter'''====
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We used colonies number 6, 7 and 8 for each promotor.
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====1 - Sequence analysis for BB pBphA1 in psB1C3 5 ( clone 4, 17 and 22 )====
 
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IMAGE
 
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==='''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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==='''B - PCB sensing system'''===
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===='''Objective : obtaining Bba_K1155002'''====
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====1 - Sequence analysis for Bba_K1155002 in clones 4, 17 and 22====
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IMAGE LOGICIEL
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The sequence is good for each clone. We obtain a new biobrick : Bba_K1155002.
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{{Team:Paris_Saclay/incl_fin}}
{{Team:Paris_Saclay/incl_fin}}

Revision as of 16:15, 16 September 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 :
[[]]
  • 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 16 : 6µL DNA Ladder
  • Well 17 to 22 : 10µL Bba_K1155004+2µl of 6X loading dye
  • Well 23 : 6µL DNA Ladder
  • Well 24 to 30 : 10µL Bba_K1155004+2µl of 6X loading dye
  • Well 31 : 6µL DNA Ladder
  • Gel : 1%
  • Bba_K1155005 :
[[]]
  • 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 :
[[]]
  • 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.  ?????????

2 - 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.


B - PCB sensing system

Objective : obtaining Bba_K1155002

1 - Sequence analysis for Bba_K1155002 in clones 4, 17 and 22

IMAGE LOGICIEL

The sequence is good for each clone. We obtain a new biobrick : Bba_K1155002.