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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<Tr><Td>PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt</Td></Tr>
<Tr><Td>PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt</Td></Tr>
</Table>
</Table>
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<b>September 4th</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</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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<Br>
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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
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<A href="https://2013.igem.org/Team:TMU-Tokyo/Notebook_experiment/2st">>>Go to next week</A>
<A href="https://2013.igem.org/Team:TMU-Tokyo/Notebook_experiment/2st">>>Go to next week</A>

Revision as of 16:37, 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

>>Go to next week