Team:TMU-Tokyo/Notebook experiment/1st

From 2013.igem.org

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<b>September 2nd</b><Br>
<b>September 2nd</b><Br>
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PCR following fragment.<Br>
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PCR following fragment. (1st PCR)<Br>
<div style="text-align:center;">
<div style="text-align:center;">
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Result<Br>
<IMG Src="https://static.igem.org/mediawiki/2013/f/f3/TMU-Tokyo_0902.jpg" width="350" hight="305"><Br>
<IMG Src="https://static.igem.org/mediawiki/2013/f/f3/TMU-Tokyo_0902.jpg" width="350" hight="305"><Br>
1: PterR1, 2: PtetR2, 3: FLP, 4: Sm, 5: araC-P,<Br>
1: PterR1, 2: PtetR2, 3: FLP, 4: Sm, 5: araC-P,<Br>
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<Br>
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<b>September 3rd</b><Br>
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Vector Remodeling with PCR. Insertion Recognition sites (Blunt).<Br>
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<div style="text-align:center;">
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<Table cellpadding="5">
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<Tr>
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<Td>Fragment</Td><Td>Km-Blunt</Td>
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</Tr>
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<Tr>
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<Td>Primer</Td><Td>Km-Blunt-N<Br>Km-Blunt-C</Td>
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</Tr>
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<Tr>
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<Td>Templete</Td><Td>pACYC177</Td>
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</Tr>
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</Table>
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</div>
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Result<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/1/19/TMU-Tokyo_0903.jpg" width="144" hight="250"><Br>
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<Br>
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<Br>
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DNA Existract following fragment from gel.<Br>
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<Table>
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<Tr><Td>PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt</Td></Tr>
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</Table>
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<Br>
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<Br>
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<b>September 4th</b><Br>
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PCR following fragment. (2nd PCR)<Br>
-
<A href="https://2013.igem.org/Team:TMU-Tokyo/Notebook_experiment/2st">>>Go to next week</A>
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<div style="text-align:center;">
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<Table cellpadding="5">
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<Tr>
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<Td>Fragment</Td><Td>Fragment 1</Td><Td>Fragment 2</Td><Td>Fragment 3</Td>
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</Tr>
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<Tr>
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<Td>Primer</Td><Td>lacA-IRR/IRL<Br>cynR-SaApL</Td><Td>araCHC<Br>araA-TcC</Td><Td>R(SaApR)-lacIN<Br>mhpA-CmC6</Td>
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</Tr>
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<Tr>
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<Td>Templete</Td><Td>PtetR<Br>FLP<Br>Sm</Td><Td>araC-P<Br>hbiF<Br>Tc</Td><Td>lacI<Br>FRT-Cm</Td>
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</Tr>
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</Table>
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</div>
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Result<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/d/d1/TMU-Tokyo_0904.jpg"><Br>
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1: Fragment 1, 2: Fragment 2, 3: Fragment 3<Br>
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<Br>
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<Br>
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<b>September 5th</b><Br>
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PCR following fragment. (2nd PCR)<Br>
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<div style="text-align:center;">
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<Table cellpadding="5">
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<Tr>
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<Td>Fragment</Td><Td>Fragment 1-A</Td><Td>Fragment 1-B</Td><Td>Fragment 2-C</Td><Td>Fragment 2-D</Td>
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</Tr>
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<Tr>
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<Td>Primer</Td><Td>CmNR-Stu<Br>FLP-R</Td><Td>FLP-L<Br>CmC</Td><Td>araC2<Br>RTcN-hbiF</Td><Td>pBAD-hbiF<Br>TcC3</Td>
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</Tr>
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<Tr>
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<Td>Templete</Td><Td>PtetR<Br>FLP</Td><Td>FLP<Br>Sm</Td><Td>araC-P<Br>hbiF</Td><Td>hbiF<Br>Tc</Td>
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</Tr>
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</Table>
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</div>
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Result<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/9/9c/TMU-Tokyo_0905.jpg"><Br>
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1: Fragment 1-A, 2: Fragment 1-B,<Br>
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3: Fragment 2-C, 4: Fragment 2-D<Br>
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<Br>
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<Br>
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Check following gel extract product.<Br>
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<Table>
 +
<Tr><Td>PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt</Td></Tr>
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</Table>
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Result<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/d/dd/TMU-Tokyo_09051.jpg"><Br>
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1: PtetR1, 2: FLP, 3: Sm, 4: araC-P, 5: hbiF,<Br>
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6: Tc, 7: lacI, 8: FRT-Cm, 9: FRT-Ap, 10: Km-Blunt<Br>
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<Br>
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<Br>
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 +
 
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<b>September 6th</b><Br>
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PCR following fragment. (2nd PCR)<Br>
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<div style="text-align:center;">
 +
<Table cellpadding="5">
 +
<Tr>
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<Td>Fragment</Td><Td>Fragment 1-A</Td><Td>Fragment 1-B</Td><Td>Fragment 2-A</Td><Td>Fragment 2-B</Td>
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</Tr>
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<Tr>
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<Td>Primer</Td><Td>CmNR-Stu<Br>FLP-R</Td><Td>FLP-L<Br>CmC</Td><Td>FLP-L<Br>CmC</Td><Td>FLP-L<Br>cynR-SaApL</Td>
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</Tr>
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<Tr>
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<Td>Templete</Td><Td>PtetR<Br>FLP</Td><Td>FLP<Br>Sm</Td><Td>Sm<Br>FLP</Td><Td>Sm<Br>FLP</Td>
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</Tr>
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</Table>
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</div>
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Result<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/a/a0/TMU-Tokyo_0906.jpg"><Br>
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1: Fragment 1-A, 2: Fragment 2-B<Br>
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(1-B and 2-A was failed in PCR)<Br>
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<Br>
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<Br>
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PCR following fragment. (2nd PCR)<Br>
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<div style="text-align:center;">
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<Table cellpadding="5">
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<Tr>
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<Td>Fragment</Td><Td>Fragment IF</Td><Td>Fragment FS</Td>
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</Tr>
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<Tr>
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<Td>Primer</Td><Td>lacA-IRR/IRL<Br>FLP-R</Td><Td>FLP-L<Br>cynR-SaApR</Td>
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</Tr>
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<Tr>
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<Td>Templete</Td><Td>PtetR<Br>FLP</Td><Td>FLP<Br>Sm</Td>
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</Tr>
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</Table>
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</div>
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<Br>
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<Br>
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<b>September 7th</b><Br>
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PCR Result (Fragment IF, FS)<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/8/89/TMU-Tokyo_0907.jpg"><Br>
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1: Fragment IF, 2: Fragment FS<Br>
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<Br>
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<Br>
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PCR following fragment. (3rd PCR)<Br>
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<div style="text-align:center;">
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<Table cellpadding="5">
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<Tr>
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<Td>Fragment</Td><Td>Fragment 1</Td><Td>Fragment 2</Td>
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</Tr>
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<Tr>
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<Td>Primer</Td><Td>lacA-IRR/IRL<Br>cynR-SaApR</Td><Td>araCHC<Br>araA-TcC</Td>
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</Tr>
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<Tr>
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<Td>Templete</Td><Td>Fragment IF<Br>Fragment FS</Td><Td>Fragment 2-B<Br>Fragment 2-C</Td>
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</Tr>
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</Table>
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</div>
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Result<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/5/52/TMU-Tokyo_09071.jpg"><Br>
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1: Fragment 1, 2: Fragment 2<Br>
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<Br>
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<Br>
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<A href="https://2013.igem.org/Team:TMU-Tokyo/Notebook_experiment/2nd">>>Go to next week</A>

Latest revision as of 21:06, 27 September 2013



Experiment > Parts Synthesis > 1st week


September 2nd
PCR following fragment. (1st PCR)
FragmentPtetR1PtetR2FLPSmaraC-P
PrimerCmNR-stu
FLP-IRR/IRL		CmCR-stu
FLP-IRR/IRL		FLP-L
FLP-R		FLP-SaApL
CmC		acaC-2
BAD-C3
TempleteIRR/IRL-GFP
(BBa_K880002)		IRR/IRL-GFP
(BBa_K880002)		pFTGTm(C2-Ap)SmpBAD/Myc-HisA
FragmenthbiFTclacIFRT-CmFRT-Ap
PrimerpBAD-HbiF
RTcN-hbiF		TcN
TcC3		141-70
FRT-lacIC		R6Kori-Xho-3R
R(FRT)2		lacI-R(FRT)2
mhpA-SaApR
TempletehbiF
(BBa_K880000)		Escherichia coli
MG1655 recA⁻
(cell suspention)		Escherichia coli
MG1655 genome		pSG76CpSG76A2
Result

1: PterR1, 2: PtetR2, 3: FLP, 4: Sm, 5: araC-P,
6: hbiF, 7: Tc, 8: LacI, 9: FRT-Cm, 10: FRT-Ap


September 3rd
Vector Remodeling with PCR. Insertion Recognition sites (Blunt).
FragmentKm-Blunt
PrimerKm-Blunt-N
Km-Blunt-C
TempletepACYC177
Result



DNA Existract following fragment from gel.
PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt


September 4th
PCR following fragment. (2nd PCR)
FragmentFragment 1Fragment 2Fragment 3
PrimerlacA-IRR/IRL
cynR-SaApL		araCHC
araA-TcC		R(SaApR)-lacIN
mhpA-CmC6
TempletePtetR
FLP
Sm		araC-P
hbiF
Tc		lacI
FRT-Cm
Result

1: Fragment 1, 2: Fragment 2, 3: Fragment 3


September 5th
PCR following fragment. (2nd PCR)
FragmentFragment 1-AFragment 1-BFragment 2-CFragment 2-D
PrimerCmNR-Stu
FLP-R		FLP-L
CmC		araC2
RTcN-hbiF		pBAD-hbiF
TcC3
TempletePtetR
FLP		FLP
Sm		araC-P
hbiF		hbiF
Tc
Result

1: Fragment 1-A, 2: Fragment 1-B,
3: Fragment 2-C, 4: Fragment 2-D


Check following gel extract product.
PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt
Result

1: PtetR1, 2: FLP, 3: Sm, 4: araC-P, 5: hbiF,
6: Tc, 7: lacI, 8: FRT-Cm, 9: FRT-Ap, 10: Km-Blunt


September 6th
PCR following fragment. (2nd PCR)
FragmentFragment 1-AFragment 1-BFragment 2-AFragment 2-B
PrimerCmNR-Stu
FLP-R		FLP-L
CmC		FLP-L
CmC		FLP-L
cynR-SaApL
TempletePtetR
FLP		FLP
Sm		Sm
FLP		Sm
FLP
Result

1: Fragment 1-A, 2: Fragment 2-B
(1-B and 2-A was failed in PCR)


PCR following fragment. (2nd PCR)
FragmentFragment IFFragment FS
PrimerlacA-IRR/IRL
FLP-R		FLP-L
cynR-SaApR
TempletePtetR
FLP		FLP
Sm


September 7th
PCR Result (Fragment IF, FS)

1: Fragment IF, 2: Fragment FS


PCR following fragment. (3rd PCR)
FragmentFragment 1Fragment 2
PrimerlacA-IRR/IRL
cynR-SaApR		araCHC
araA-TcC
TempleteFragment IF
Fragment FS		Fragment 2-B
Fragment 2-C
Result

1: Fragment 1, 2: Fragment 2


>>Go to next week

Retrieved from "http://2013.igem.org/Team:TMU-Tokyo/Notebook_experiment/1st"