Team:Paris Saclay/Notebook/August/6

From 2013.igem.org

(Difference between revisions)
Line 7: Line 7:
==='''A - Aerobic/Anaerobic regulation system'''===
==='''A - Aerobic/Anaerobic regulation system'''===
-
===='''Obtaining activator FNR promoter (narK, nirB, narG)'''====
+
===='''Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006'''====
 +
 
 +
====1 - Electrophoresis to check the Colony PCR products : Bba_K1155004, Bba_K1155005, Bba_K1155006====
-
====1 - Gel electrophoresis of the colony PCR products ( made the 08/05/2013)on 25 colonies====
 
XiaoJing, Damir, Anaïs
XiaoJing, Damir, Anaïs
-
* NarK promoter in psB1C3
+
* Bba_K1155004 :
{|
{|
-
| style="width:350px;border:1px solid black;" | IMAGE
+
| style="width:350px;border:1px solid black;" |[[]]
| style="width:350px;border:1px solid black;vertical-align:top;" |
| style="width:350px;border:1px solid black;vertical-align:top;" |
-
*Wells 1 to 25 : 10µl sample
+
* Well 1 : 6µL DNA Ladder
-
*Well between 12 and 13 : 6µl gene ruler
+
* Well 2 to 7 : 10µL Bba_K1155004+2µl of 6X loading dye
-
*Gel : 1.5%
+
* 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%
|}
|}
-
Expected size : 500 bp
+
*Bba_K1155005 :
-
 
+
-
We obtained fragments of the right size. Our backbone contains the Biobrick.
+
-
 
+
-
*NirB promoter in psB1C3
+
{|
{|
-
| style="width:350px;border:1px solid black;" | IMAGE
+
| style="width:350px;border:1px solid black;" |[[]]
| style="width:350px;border:1px solid black;vertical-align:top;" |
| style="width:350px;border:1px solid black;vertical-align:top;" |
-
*Before well 1 : 6µl gene ruler
+
* Well 1 : 6µL DNA Ladder
-
*Wells 1 to 6 : 10µl sample
+
* Well 2 to 7 : 10µL Bba_K1155005+2µl of 6X loading dye
-
*Between 6 and 7: 6µl gene ruler
+
* Well 8 : 6µL DNA Ladder
-
*Wells 7 to 12 : 10µl sample
+
* Well 9 to 14 : 10µL Bba_K1155005+2µl of 6X loading dye
-
*After well 12 :6µl gene ruler
+
* Well 15 : 6µL DNA Ladder
-
*Before well 13  : 6µl gene ruler
+
* Well 16 : 6µL DNA Ladder
-
*Wells 13 to 18 : 10µl sample
+
* Well 17 to 22 : 10µL Bba_K1155005+2µl of 6X loading dye
-
*Between 18 and 19: 6µl gene ruler
+
* Well 23 : 6µL DNA Ladder
-
*Wells 19 to 25 : 10µl sample
+
* Well 24 to 30 : 10µL Bba_K1155005+2µl of 6X loading dye
-
*After well 25 :6µl gene ruler
+
* Well 31 : 6µL DNA Ladder
-
*Gel : 1.5%
+
* Gel : 1%
|}
|}
-
Expected size : 500 bp
+
*Bba_K1155006 :
-
 
+
-
We obtained fragments of the right size. Our backbone contains the Biobrick.
+
-
 
+
-
*NarG promoter in psB1C3
+
{|
{|
-
| style="width:350px;border:1px solid black;" | IMAGE
+
| style="width:350px;border:1px solid black;" |[[]]
| style="width:350px;border:1px solid black;vertical-align:top;" |
| style="width:350px;border:1px solid black;vertical-align:top;" |
-
*Before well 1 : 6µl gene ruler
+
* Well 1 to 12 : 10µL Bba_K1155006+2µl of 6X loading dye
-
*Wells 1 to 6 : 10µl sample
+
* Well 13 : 6µL DNA Ladder
-
*Between 6 and 7: 6µl gene ruler
+
* Well 14 to 26 : 10µL Bba_K1155006+2µl of 6X loading dye
-
*Wells 7 to 12 : 10µl sample
+
* Gel : 1%
-
*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
+
Expected size :
 +
* NarK, NarG, NirB : 500 bp
-
We obtained fragments of the right size. Our backbone contains the Biobrick.
+
{|
 +
| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
 +
We obtain fragments at the good size for all the colony. ENSUITE ?????????
 +
|}
====2 - Samples culture====
====2 - Samples culture====

Revision as of 15:49, 16 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. ENSUITE ?????????

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

Retrieved from "http://2013.igem.org/Team:Paris_Saclay/Notebook/August/6"