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Notebook : August 19

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155007

1 - Sequence analysis of BBa_K1155007

N'Guyen

Our sequence analysis was good. We obtain our biobrick BBa_K1155007.

Objective : characterize BBa_K1155000, BBa_K1155004, BBa_K155005, BBa_K1155006

1 - EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term

Nadia

Protocol : EtOH precipitation

We used 20µL of DNA.

Nanodrop :

RBS-LacZ-Term : 38.4ng/µL
RBS-AmilCP-Term : 28.7ng/µL

The precipitation was good. We will make ligations.

2 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3

XiaoJing

Used quantities :

NirB and RBS-LacZ-Term :
- NirB : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL

NirB and RBS-AmilCP-Term :
- NirB : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL

NarG and RBS-LacZ-Term :
- NarG : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL

NarG and RBS-AmilCP-Term :
- NarG : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL

NarK and RBS-LacZ-Term :
- Nar K : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL

NarK and RBS-AmilCP-Term :
- NarK : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL

3 - Transformation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α

XiaoJing

Protocol : Bacterial transformation

4- Make competent cells of strain MG1655Z1 Δfnr::Km

XiaoJing

Protocol : Preparation of super competent cells

5- Transformation of MG1655Z1 Δfnr::Km

XiaoJing

We transformed pcp20 plasmid into strain MG1655Z1 Δfnr::Km. Then we spread in LB and ampicillin antibiotic plate. We need to incubate it at 30°C for two days.

Protocol : Bacterial transformation

A - Aerobic/Anaerobic regulation system / B - PCB sensor system

Objective : obtaining FNR and BphR2 proteins

1 - Electrophoresis gel of PCR products : BphR2 Part I

Damir

Well 1 : 6µL DNA Ladder
Well 2 : 40µL BphR2 part I + 8µl of 6X loading dye
Gel : 1%

Expected size :

BphR2 Part I : 178 kb

We obtained a frangment at the right size. We can purify it.

2 - Gel purification of PCR products : BphR2 Part I

Damir

Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]

Nanodrop :

BphR2 Part I: 57.2ng/µL

We lost our fragment BphR2 Part I. We will make the PCR again.

Lab work, Modeling, Human practices

Meeting with Evry

We spent our afternoon with Evry team. Each team presented its lab, modeling and human practices project. We learned informations to improve our human practices project. We discussed about safety and gave them some advices about the pills they would make.

Previous day

Back to calendar

Next day

@@ Line 7: / Line 7: @@
 ==='''A - Aerobic/Anaerobic regulation system'''===
-===='''Objective : to obtain ... '''====
+===='''Objective : obtaining BBa_K1155007'''====
-=====1 - Gel purification of the digestion of Bba_K1155003 and Bba_K1155007 by XBaI and PstI=====
+====1 - Sequence analysis of BBa_K1155007====
-Nadia
-Protocol : [[Team:Paris_Saclay/Protocols/Gel purification|Gel purification]]
+N'Guyen
-Nanodrop :
+{|
-* RBS-LacZ-Term : 59.6ng/µL
+| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
-* RBS-AmilCP-Term : 21.3ng/µL
+Our sequence analysis was good. We obtain our biobrick BBa_K1155007.
+|}
- Concentrations are very low so we try to precipitate the DNA again.
+===='''Objective : characterize BBa_K1155000, BBa_K1155004, BBa_K155005, BBa_K1155006 '''====
-=====2 - EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term=====
+====1 - EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term====
-Nadia, XiaoJing
-Protocol : [[Team:Paris_Saclay/Protocols/EtOH precipitation|EtOH precipitation]]
+Nadia
+Protocol : [[Team:Paris_Saclay/ethanol|EtOH precipitation]]
 We used 20µL of DNA.
-==='''A - Aerobic/Anaerobic regulation system - B - PCB sensor system'''===
+Nanodrop :
+* RBS-LacZ-Term : 38.4ng/µL
+* RBS-AmilCP-Term : 28.7ng/µL
-===='''Objective : to obtain ... '''====
+{|
+| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
+The precipitation was good. We will make ligations.
+|}
+====2 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3====
+XiaoJing
+Used quantities :
+* NirB and RBS-LacZ-Term :
+** NirB : 3µL
+** RBS-LacZ-Term : 1µL
+** Buffer : 2µL
+** H2O : 13µL
+** Ligase : 1µL
+* NirB and RBS-AmilCP-Term :
+** NirB : 3µL
+** RBS-AmilCP-Term : 1µL
+** Buffer : 2µL
+** H2O : 13µL
+** Ligase : 1µL
+* NarG and RBS-LacZ-Term :
+** NarG : 3µL
+** RBS-LacZ-Term : 1µL
+** Buffer : 2µL
+** H2O : 13µL
+** Ligase : 1µL
+* NarG and RBS-AmilCP-Term :
+** NarG : 3µL
+** RBS-AmilCP-Term : 1µL
+** Buffer : 2µL
+** H2O : 13µL
+** Ligase : 1µL
+* NarK and RBS-LacZ-Term :
+** Nar K : 3µL
+** RBS-LacZ-Term : 1µL
+** Buffer : 2µL
+** H2O : 13µL
+** Ligase : 1µL
+* NarK and RBS-AmilCP-Term :
+** NarK : 3µL
+** RBS-AmilCP-Term : 1µL
+** Buffer : 2µL
+** H2O : 13µL
+** Ligase : 1µL
+====3 - Transformation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α====
+XiaoJing
+Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
+====4- Make competent cells of strain MG1655Z1 Δfnr::Km====
+XiaoJing
+Protocol : [[Team:Paris_Saclay/preparation|Preparation of super competent cells]]
+====5- Transformation of MG1655Z1 Δfnr::Km====
+XiaoJing
+We transformed pcp20 plasmid into strain MG1655Z1 Δfnr::Km. Then we spread in LB and ampicillin antibiotic plate. We need to incubate it at 30°C for two days.
+Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
+==='''A - Aerobic/Anaerobic regulation system / B - PCB sensor system'''===
+===='''Objective : obtaining FNR and BphR2 proteins'''====
+====1 - Electrophoresis gel of PCR products : BphR2 Part I====
-=====1 - Electrophoresis gel of BphR2 produce by PCR=====
 Damir
 {|
-| style="width:250px;border:1px solid black;" | [[File:PSgel1908.jpg]]
+| style="width:250px;border:1px solid black;" | [[File:PSgel11908.jpg]]
 | style="width:350px;border:1px solid black;vertical-align:top;" |
 *Well 1 : 6µL DNA Ladder
-*Well 2 : 5µL DNA + 1µL Dyes
+*Well 2 : 40µL BphR2 part I + 8µl of 6X loading dye
 *Gel : 1%
 |}
- We obtain the BphR2 by PCR.
+Expected size :
+* BphR2 Part I : 178 kb
+{|
+| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
+We obtained a frangment at the right size. We can purify it.
+|}
+====2 - Gel purification of PCR products : BphR2 Part I ====
-=====2 - Gel purification of BphR2 =====
 Damir
-Protocol : [[Team:Paris_Saclay/Protocols/Gel purification|Gel purification]]
+Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]
 Nanodrop :
-* BphR2 : 57.2ng/µL
+* BphR2 Part I: 57.2ng/µL
+ {|
+| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
+We lost our fragment BphR2 Part I. We will make the PCR again.
+|}
+=='''Lab work, Modeling, Human practices'''==
+=='''Meeting with Evry'''==
+[[File:PSevry2.jpg]]
+We spent our afternoon with Evry team. Each team presented its lab, modeling and human practices project. We learned informations to improve our human practices project. We discussed about safety and gave them some advices about the pills they would make.
+{| border="1" align="center"
+|[[Team:Paris Saclay/Notebook/August/14|<big>Previous day</big>]]
+|[[Team:Paris_Saclay/Notebook|<big>Back to calendar</big>]]
+|[[Team:Paris Saclay/Notebook/August/20|<big>Next day</big>]]
+|}
 {{Team:Paris_Saclay/incl_fin}}

Team:Paris Saclay/Notebook/August/19

From 2013.igem.org

Latest revision as of 01:29, 5 October 2013

Contents

Notebook : August 19

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155007

1 - Sequence analysis of BBa_K1155007

Objective : characterize BBa_K1155000, BBa_K1155004, BBa_K155005, BBa_K1155006

1 - EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term

2 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3

3 - Transformation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in pSB1C3 in DH5α

4- Make competent cells of strain MG1655Z1 Δfnr::Km

5- Transformation of MG1655Z1 Δfnr::Km

A - Aerobic/Anaerobic regulation system / B - PCB sensor system

Objective : obtaining FNR and BphR2 proteins

1 - Electrophoresis gel of PCR products : BphR2 Part I

2 - Gel purification of PCR products : BphR2 Part I

Lab work, Modeling, Human practices

Meeting with Evry