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. ?????????
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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.
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We used colonies number 6, 7 and 8 for each promotor.
We used colonies number 6, 7 and 8 for each promotor.
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===='''Objective : obtaining Bba_K1155007'''====
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====1 - Electroelution of Bba_I732017 digested by ecoRI/SpeI====
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Nadia
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Protocol : [[Team:Paris_Saclay/Protocols/Electroelution|Electroelution]]
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The electroelution was good. We will ligate RBS-LacZ with Term and PSB1C3.
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==='''B - PCB sensing system'''===
==='''B - PCB sensing system'''===

Revision as of 17:44, 20 September 2013

Navigation Home Team Project Labwork Notebook Human practices

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. We will make a culture of DH5α with Bba_K1155004, Bba_K1155005 and Bba_K1155006. We will also sequence our plasmids.

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.

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.

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.


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