Team:KAIT Japan/Notebook

From 2012.igem.org

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<P>[[File:KAIT_Japan2012_logo.png|left|link=Team:KAIT_Japan]][[File:KAIT_Japan2012_Sub.png|right|link=http://www.kait.jp/index2.php]][[File:Kaitjapan_iGEM_official.logo.png‎|right|link=http://ung.igem.org/Main_Page]]
<P>[[File:KAIT_Japan2012_logo.png|left|link=Team:KAIT_Japan]][[File:KAIT_Japan2012_Sub.png|right|link=http://www.kait.jp/index2.php]][[File:Kaitjapan_iGEM_official.logo.png‎|right|link=http://ung.igem.org/Main_Page]]
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</P><br>
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{|style="color:#000;background-color:#FFF;" width="100%" align="center"  
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{|style="background:#99F; margin:0.1em"
!align="center"|[[File:Kaitjapan.home.png|link=Team:KAIT_Japan]]
!align="center"|[[File:Kaitjapan.home.png|link=Team:KAIT_Japan]]
[[Team:KAIT_Japan|Home]]
[[Team:KAIT_Japan|Home]]
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{|style="background:#99F; margin:0.1em"
!align="center"|[[File:Kaitjapan_project.png|link=Team:KAIT_Japan/Project]]
!align="center"|[[File:Kaitjapan_project.png|link=Team:KAIT_Japan/Project]]
[[Team:KAIT_Japan/Project|Project]]
[[Team:KAIT_Japan/Project|Project]]
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{|style="background:#99F; margin:0.1em"
!align="center"|[[File:Kaitjapan_parts.png|link=Team:KAIT_Japan/Parts]]
!align="center"|[[File:Kaitjapan_parts.png|link=Team:KAIT_Japan/Parts]]
[[Team:KAIT_Japan/Parts|Parts]]
[[Team:KAIT_Japan/Parts|Parts]]
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{|style="background:#99F; margin:0.1em"
!align="center"|[[File:Kaitjapan_protocol.png|link=Team:KAIT_Japan/Protocol]]
!align="center"|[[File:Kaitjapan_protocol.png|link=Team:KAIT_Japan/Protocol]]
[[Team:KAIT_Japan/Protocol|Protocol]]
[[Team:KAIT_Japan/Protocol|Protocol]]
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{|style="background:#99F; margin:0.1em"
!align="center"|[[File:Kaitjapan_notebook.png|link=Team:KAIT_Japan/Notebook]]
!align="center"|[[File:Kaitjapan_notebook.png|link=Team:KAIT_Japan/Notebook]]
[[Team:KAIT_Japan/Notebook|Notebook]]
[[Team:KAIT_Japan/Notebook|Notebook]]
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{|style="background:#99F; margin:0.1em"
!align="center"|[[File:Kaitjapan_results.png|link=Team:KAIT_Japan/Results]]
!align="center"|[[File:Kaitjapan_results.png|link=Team:KAIT_Japan/Results]]
[[Team:KAIT_Japan/Results|Results]]
[[Team:KAIT_Japan/Results|Results]]
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{|style="background:#99F; margin:0.1em"
!align="center"|[[File:Kaitjapan_safety.png|link=Team:KAIT_Japan/Safety]]
!align="center"|[[File:Kaitjapan_safety.png|link=Team:KAIT_Japan/Safety]]
[[Team:KAIT_Japan/Safety|Safety]]
[[Team:KAIT_Japan/Safety|Safety]]
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{|style="background:#99F; margin:0.1em"
!align="center"|[[File:Kaitjapan_human_practice.png|link=Team:KAIT_Japan/Human_Practice]]
!align="center"|[[File:Kaitjapan_human_practice.png|link=Team:KAIT_Japan/Human_Practice]]
[[Team:KAIT_Japan/Human_Practice|Human Practice]]
[[Team:KAIT_Japan/Human_Practice|Human Practice]]
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{|style="background:#99F; margin:0.1em"
!align="center"|[[File:Kaitjapan_team.png|link=Team:KAIT_Japan/Team]]
!align="center"|[[File:Kaitjapan_team.png|link=Team:KAIT_Japan/Team]]
[[Team:KAIT_Japan/Team|Team]]
[[Team:KAIT_Japan/Team|Team]]
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__NOTOC__
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='''Notebook'''=
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:We made an experiment every day. It was process of trial and error.
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=='''Creating parts of Methylation and Azurin'''==
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:<big>'''Date:8/9'''</big> PCR
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We are creating now...<br>
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:<big>'''Date:8/10'''</big> Electrophoresis
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=Creating parts of Tar methylation region.=
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==Date:8/9==
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===Colony PCR===
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Reagent
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*TaKaRa Ex Taq(5units/μL) 0.5μL
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*10×Ex Taq buffer 10μL
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*dNTP Mixture(2.5Meach) 8μL
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*Primer F(10μM) 4μL
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*Primer R(10μM) 4μL
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*Template(E.coli DH5α)
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'''→Reflection:We Took E.coli too many.You should take less E.coli.'''<br>
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Conditions of the thermal cycler
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#95°C(5min)
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#94°C(30sec)
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#61°C(30sec)
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#71°C(40sec)
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#72°C(1min)
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#4°C(Save)
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#*2~4:35cycle'''→Reflection:The number of cycles was less.So,We increased The number of cycles in 8/11.(50 cycles) '''
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----
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:<big>'''Date:8/13'''</big> Electrophoresis and The refinement of DNA,PCR
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==Date:8/11==
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:<big>'''Date:8/14'''</big> Transformation
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===The purified DNA===
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#Electrophoresis
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#*Marker:pigment(buffer) 1μL,DNA molecule 2μL,TE buffer 3μL
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#*Sample:pigment(buffer) 1μL,sample 5μL
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#*Gel concentration:1.2%,Migration time:30min
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'''→Reflection:Band was less.'''
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#Storage
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===PCR Product===
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:<big>'''Date:8/16'''</big> Blue white selection and Miniprep,Colony PCR
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#Electrophoresis
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#*The gel check and cut
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#DNA purification
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#Confirmation of electrophoresis
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#PCR
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#*50cycle
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#Storage
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----
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:<big>'''Date:8/17'''</big> Electrophoresis and Colony PCR
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==Date:8/13==
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:<big>'''Date:8/18'''</big> PCR and Electrophoresis
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===Confirmed of electrophoresis by PCR product and Ligation of the TA vector===
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#Electrophoresis
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#*Gel concentration:1.2%,Migration time:30min
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#*Marker:Flash Gel 5μL
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#*Sample:pigment 1μL,sample 5μL
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#Check and Colony PCR
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#Add to TA vector
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#*PCR product 2μL
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#*pMD20-Tvector 1μL
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#*D<sub>2</sub>W 2μL
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#*Ligation Mighty Mix 5μL
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#Heat insulation(16°C,30min)
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#Storage(-20°C)
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----
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:<big>'''Date:8/20'''</big> Extraction and the refinement of DNA
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==Date:8/14==
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:<big>'''Date:8/21'''</big> PCR and Electrophoresis,Transformation
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===Transformation===
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#Put competent cells on ice(10~15min)
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:<big>'''Date:8/22'''</big> Electrophoresis
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#Add Ligation reaction solution(10μL) and tapping
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#On the ice(30min)[Transformation]
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:<big>'''Date:8/23'''</big> PCR
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#Add LB medium(0.7mL)
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#Incubate(60min,37°C)
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:<big>'''Date:8/25'''</big> Miniprep and Restriction enzyme processing
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#Add X-gal(40μL) and ampicillin(10μL)[200μg/mL] on LB agar medium(IPTG)
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#Add one incubated(100μL)
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:<big>'''Date:8/27'''</big> Electrophoresis
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#Cultivation(overnight)
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 +
:<big>'''Date:8/28'''</big> Electrophoresis and The refinement of DNA,PCR
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 +
:<big>'''Date:8/29'''</big> Electrophoresis
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 +
:<big>'''Date:8/30'''</big> Electrophoresis and The refinement of DNA
 +
 
 +
:<big>'''Date:9/1'''</big> Extraction of DNA
 +
 
 +
:<big>'''Date:9/2'''</big> Redesign of the primer
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 +
:<big>'''Date:9/3'''</big> PCR and Restriction enzyme processing,Electrophoresis
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 +
:<big>'''Date:9/5'''</big> Miniprep
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 +
:<big>'''Date:9/6'''</big> Electrophoresis
 +
 
 +
:<big>'''Date:9/10'''</big> Colony PCR
 +
 
 +
:<big>'''Date:9/12'''</big> Miniprep
 +
 
 +
:<big>'''Date:9/14'''</big> PCR
 +
 
 +
:<big>'''Date:9/15'''</big> Restriction enzyme processing
 +
 
 +
:<big>'''Date:9/16'''</big> PCR
 +
 
 +
:<big>'''Date:9/17'''</big> PCR
 +
 
 +
:<big>'''Date:9/19'''</big> PCR and Restriction enzyme processing
 +
 
 +
:<big>'''Date:9/20'''</big> Ligation
 +
 
 +
:<big>'''Date:9/22'''</big> Transformation
 +
 
 +
:<big>'''Date:9/23'''</big> PCR and Electrophoresis
 +
 
 +
:<big>'''Date:9/24'''</big> Ligation
 +
 
 +
:<big>'''Date:9/25'''</big> Transformation
 +
 
 +
:<big>'''Date:9/27'''</big> Miniprep and Electrophoresis
 +
 
 +
=='''Creating parts of Tar'''==
 +
:<big>'''Date:9/13'''</big> PCR and Colony PCR
 +
 
 +
:<big>'''Date:9/14'''</big> Electrophoretic confirmation
 +
 
 +
:<big>'''Date:9/15'''</big> Restriction enzyme processing
 +
 
 +
:<big>'''Date:9/16'''</big> Electrophoresis of the restriction enzyme processing product and Restriction enzyme processing
 +
 
 +
:<big>'''Date:9/17'''</big> Refinement of DNA and Ligation,Transformation
 +
 
 +
:<big>'''Date:9/18'''</big> Making of a master plate and Preculture,Recultivation
 +
 
 +
:<big>'''Date:9/19'''</big> Colony PCR and Electrophoresis
 +
 
 +
:<big>'''Date:9/20'''</big> Ligation and Transformation
 +
 
 +
:<big>'''Date:9/21'''</big> Inoculation
 +
 
 +
:<big>'''Date:9/22'''</big> Making of a master plate and Colony PCR,Electrophoresis,Preculture
 +
 
 +
:<big>'''Date:9/23'''</big> Electrophoretic confirmation and Electrophoresis of the PCR
 +
 
 +
:<big>'''Date:9/24'''</big> Miniprep and Electrophoresis
 +
 
 +
:<big>'''Date:9/25'''</big> PCR of the miniprep product and Electrophoresis
 +
 
 +
=='''Creating parts of EnvZ'''==
 +
:<big>'''Date:9/10'''</big> Design of the primer and PCR,Elecrtophoresis,Miniprep
 +
 
 +
:<big>'''Date:9/13'''</big> PCR and Elecrtophoresis
 +
 
 +
:<big>'''Date:9/18'''</big> Design of the primer
 +
 
 +
:<big>'''Date:9/19'''</big> PCR and Elecrtophoresis
 +
 
 +
:<big>'''Date:9/20'''</big> Restriction enzyme processing
 +
 
 +
:<big>'''Date:9/21'''</big> Ligation and PCR,Elecrtophoresis
 +
 
 +
:<big>'''Date:9/23'''</big> PCR
 +
 
 +
:<big>'''Date:9/24'''</big> The refinement of DNA and Restriction enzyme processing,Ligation
 +
 
 +
:<big>'''Date:9/25'''</big> Transformation
 +
 
 +
:<big>'''Date:9/26'''</big> Liquid culture
 +
 
 +
|}

Latest revision as of 22:12, 26 September 2012

KAIT Japan2012 logo.png
KAIT Japan2012 Sub.png
Kaitjapan iGEM official.logo.png


Kaitjapan.home.png

Home

Kaitjapan project.png

Project

Kaitjapan parts.png

Parts

Kaitjapan protocol.png

Protocol

Kaitjapan notebook.png

Notebook

Kaitjapan results.png

Results

Kaitjapan safety.png

Safety

Kaitjapan human practice.png

Human Practice

Kaitjapan team.png

Team

Notebook

We made an experiment every day. It was process of trial and error.

Creating parts of Methylation and Azurin

Date:8/9 PCR
Date:8/10 Electrophoresis
Date:8/13 Electrophoresis and The refinement of DNA,PCR
Date:8/14 Transformation
Date:8/16 Blue white selection and Miniprep,Colony PCR
Date:8/17 Electrophoresis and Colony PCR
Date:8/18 PCR and Electrophoresis
Date:8/20 Extraction and the refinement of DNA
Date:8/21 PCR and Electrophoresis,Transformation
Date:8/22 Electrophoresis
Date:8/23 PCR
Date:8/25 Miniprep and Restriction enzyme processing
Date:8/27 Electrophoresis
Date:8/28 Electrophoresis and The refinement of DNA,PCR
Date:8/29 Electrophoresis
Date:8/30 Electrophoresis and The refinement of DNA
Date:9/1 Extraction of DNA
Date:9/2 Redesign of the primer
Date:9/3 PCR and Restriction enzyme processing,Electrophoresis
Date:9/5 Miniprep
Date:9/6 Electrophoresis
Date:9/10 Colony PCR
Date:9/12 Miniprep
Date:9/14 PCR
Date:9/15 Restriction enzyme processing
Date:9/16 PCR
Date:9/17 PCR
Date:9/19 PCR and Restriction enzyme processing
Date:9/20 Ligation
Date:9/22 Transformation
Date:9/23 PCR and Electrophoresis
Date:9/24 Ligation
Date:9/25 Transformation
Date:9/27 Miniprep and Electrophoresis

Creating parts of Tar

Date:9/13 PCR and Colony PCR
Date:9/14 Electrophoretic confirmation
Date:9/15 Restriction enzyme processing
Date:9/16 Electrophoresis of the restriction enzyme processing product and Restriction enzyme processing
Date:9/17 Refinement of DNA and Ligation,Transformation
Date:9/18 Making of a master plate and Preculture,Recultivation
Date:9/19 Colony PCR and Electrophoresis
Date:9/20 Ligation and Transformation
Date:9/21 Inoculation
Date:9/22 Making of a master plate and Colony PCR,Electrophoresis,Preculture
Date:9/23 Electrophoretic confirmation and Electrophoresis of the PCR
Date:9/24 Miniprep and Electrophoresis
Date:9/25 PCR of the miniprep product and Electrophoresis

Creating parts of EnvZ

Date:9/10 Design of the primer and PCR,Elecrtophoresis,Miniprep
Date:9/13 PCR and Elecrtophoresis
Date:9/18 Design of the primer
Date:9/19 PCR and Elecrtophoresis
Date:9/20 Restriction enzyme processing
Date:9/21 Ligation and PCR,Elecrtophoresis
Date:9/23 PCR
Date:9/24 The refinement of DNA and Restriction enzyme processing,Ligation
Date:9/25 Transformation
Date:9/26 Liquid culture
Retrieved from "http://2012.igem.org/Team:KAIT_Japan/Notebook"