Team:Paris Saclay/Notebook/August/6

From 2013.igem.org

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(→Objective : obtaining biobricks in PSB3K3)
(→1 - Electrophoresis to check the PCR Colonies products : BBa_K1155004, BBa_K1155005, BBa_K1155006)
 
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===='''Objective : obtaining BBa_K1155004, BBa_K1155005, BBa_K1155006'''====
===='''Objective : obtaining BBa_K1155004, BBa_K1155005, BBa_K1155006'''====
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====1 - Electrophoresis to check the Colony PCR products : BBa_K1155004, BBa_K1155005, BBa_K1155006====
+
====1 - Electrophoresis to check the PCR Colonies products : BBa_K1155004, BBa_K1155005, BBa_K1155006====
XiaoJing, Damir, Anaïs
XiaoJing, Damir, Anaïs
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| style="width:350px;border:1px solid black;vertical-align:top;" |
| style="width:350px;border:1px solid black;vertical-align:top;" |
* Well 1 : 6µL DNA Ladder
* 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 2 to 7 : 10µL BBa_K1155004 + 2µl of 6X loading dye
* Well 8 : 6µL DNA Ladder
* Well 8 : 6µL DNA Ladder
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* Well 9 to 15 : 10µL BBa_K1155004+2µl of 6X loading dye
+
* Well 9 to 15 : 10µL BBa_K1155004 + 2µl of 6X loading dye
* Well 16 : 6µL DNA Ladder
* Well 16 : 6µL DNA Ladder
* Well 1 : 6µL DNA Ladder
* Well 1 : 6µL DNA Ladder
-
* Well 2 to 7 : 10µL BBa_K1155004+2µl of 6X loading dye
+
* Well 2 to 7 : 10µL BBa_K1155004 + 2µl of 6X loading dye
* Well 8 : 6µL DNA Ladder
* Well 8 : 6µL DNA Ladder
-
* 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
* Gel : 1%
* Gel : 1%
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| style="width:350px;border:1px solid black;vertical-align:top;" |
| style="width:350px;border:1px solid black;vertical-align:top;" |
* Well 1 : 6µL DNA Ladder
* Well 1 : 6µL DNA Ladder
-
* Well 2 to 7 : 10µL BBa_K1155005+2µl of 6X loading dye
+
* Well 2 to 7 : 10µL BBa_K1155005 + 2µl of 6X loading dye
* Well 8 : 6µL DNA Ladder
* Well 8 : 6µL DNA Ladder
-
* Well 9 to 14 : 10µL BBa_K1155005+2µl of 6X loading dye
+
* Well 9 to 14 : 10µL BBa_K1155005 + 2µl of 6X loading dye
* Well 15 : 6µL DNA Ladder
* Well 15 : 6µL DNA Ladder
* Well 1 : 6µL DNA Ladder
* Well 1 : 6µL DNA Ladder
-
* Well 2 to 7 : 10µL BBa_K1155005+2µl of 6X loading dye
+
* Well 2 to 7 : 10µL BBa_K1155005 + 2µl of 6X loading dye
* Well 8 : 6µL DNA Ladder
* Well 8 : 6µL DNA Ladder
-
* Well 9 to 15 : 10µL BBa_K1155005+2µl of 6X loading dye
+
* Well 9 to 15 : 10µL BBa_K1155005 + 2µl of 6X loading dye
* Well 16 : 6µL DNA Ladder
* Well 16 : 6µL DNA Ladder
* Gel : 1%
* Gel : 1%
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{|
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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.
+
We obtained fragments of the good size for all the colonies. We will make a culture for DH5α strain containing BBa_K1155004, BBa_K1155005 and BBa_K1155006. We will also sequence our plasmids.
|}
|}
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====2 - Liquid culture of DH5α with BBa_K1155004, BBa_K1155005 and BBa_K1155006 ====
+
====2 - Liquid culture of DH5αstrain containing BBa_K1155004, BBa_K1155005 and BBa_K1155006 ====
Xiaoing, Anaïs
Xiaoing, Anaïs
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Used quantities :  
Used quantities :  
* LB : 5mL  
* LB : 5mL  
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* Chloramphénicol (1000X, 20µg/mL) : 5µL
+
* Chloramphenicol (1000X, 20µg/mL) : 5µL
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* BBa_K1155004, BBa_K1155005 and BBa_K1155006 : 25µL
+
* Strain containing BBa_K1155004, BBa_K1155005 and BBa_K1155006 : 25µL
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We let the incubation over night at 37°C at 180 RPM.
+
We incubate the cultures over night at 37°C with  agigation at 180 RPM.
-
We used colonies number 6, 7 and 8 for each promotor.
+
We picked colonies number 6, 7 and 8 for each promoter.
===='''Objective : obtaining BBa_K1155007'''====
===='''Objective : obtaining BBa_K1155007'''====

Latest revision as of 01:25, 5 October 2013

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Contents

Notebook : August 6

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155004, BBa_K1155005, BBa_K1155006

1 - Electrophoresis to check the PCR Colonies 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 15 : 10µL BBa_K1155004 + 2µl of 6X loading dye
  • Well 16 : 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 14 : 10µL BBa_K1155004 + 2µl of 6X loading dye
  • Well 15 : 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 1 : 6µL DNA Ladder
  • Well 2 to 7 : 10µL BBa_K1155005 + 2µl of 6X loading dye
  • Well 8 : 6µL DNA Ladder
  • Well 9 to 15 : 10µL BBa_K1155005 + 2µl of 6X loading dye
  • Well 16 : 6µL DNA Ladder
  • Gel : 1%
  • BBa_K1155006 :
Psgel50608.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 obtained fragments of the good size for all the colonies. We will make a culture for DH5α strain containing BBa_K1155004, BBa_K1155005 and BBa_K1155006. We will also sequence our plasmids.

2 - Liquid culture of DH5αstrain containing BBa_K1155004, BBa_K1155005 and BBa_K1155006

Xiaoing, Anaïs

Used quantities :

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

We incubate the cultures over night at 37°C with agigation at 180 RPM.

We picked colonies number 6, 7 and 8 for each promoter.

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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