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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 :

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 :

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 :

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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@@ Line 7: / Line 7: @@
 ==='''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 PCR Colonies 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
-* NarK promoter in psB1C3
+* BBa_K1155004 :
 {|
-| style="width:350px;border:1px solid black;" | IMAGE
+| style="width:350px;border:1px solid black;" |[[File:Psgel10608.jpg| 400px]][[File:Psgel20608.jpg| 400px]]
 | 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 15 : 10µL BBa_K1155004 + 2µl of 6X loading dye
+* Well 16 : 6µL DNA Ladder
-Expected size : 500 bp
- We obtained fragments of the right size. Our backbone contains the Biobrick.
+* 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%
+|}
-*NirB promoter in psB1C3
+*BBa_K1155005 :
 {|
-| style="width:350px;border:1px solid black;" | IMAGE
+| style="width:350px;border:1px solid black;" |[[File:Psgel30608.jpg| 400px]][[File:Psgel40608.jpg| 400px]]
 | 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
-*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
- We obtained fragments of the right size. Our backbone contains the Biobrick.
+* 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%
+|}
-*NarG promoter in psB1C3
+*BBa_K1155006 :
 {|
-| style="width:350px;border:1px solid black;" | IMAGE
+| style="width:350px;border:1px solid black;" |[[File:Psgel50608.jpg| 400px]]
 | 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 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 ====
-====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)
+Used quantities :
-Incubator over night to 310,15K at 180 RPM
+* 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 : [[Team:Paris_Saclay/electro|Electroelution]]
+{|
+| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
+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 : [[Team:Paris_Saclay/extraction|Low copy plamid extraction]]
 ==='''B - PCB sensing system'''===
-===='''Obtaining the BphA1 promoter'''====
+===='''Objective : obtaining BBa_K1155002'''====
-====1 -
+====1 - Sequence analysis for BBa_K1155002 in clones 4, 17 and 22====
+{|
+| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
+The sequence is good for each clone. We obtain a new biobrick : BBa_K1155002.
+|}
+{| border="1" align="center"
+|[[Team:Paris Saclay/Notebook/August/5|<big>Previous day</big>]]
+|[[Team:Paris_Saclay/Notebook|<big>Back to calendar</big>]]
+|[[Team:Paris Saclay/Notebook/August/7|<big>Next day</big>]]
+|}
 {{Team:Paris_Saclay/incl_fin}}

Team:Paris Saclay/Notebook/August/6