Team:TMU-Tokyo/Notebook experiment/3rd

From 2013.igem.org

(Difference between revisions)
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<b>September 17th</b><Br>
<b>September 17th</b><Br>
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PCR result.<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/7/7c/TMU-Tokyo_0917.jpg"><Br>
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1-2: Fragment 1, 3-4: Fragment 2,<Br>
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5-6: Fragment 3 - Cm, 8-10: Fragment 3 - Ap<Br>
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<Br>
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<Br>
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Electro Poration<Br>
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<Table><Tr><Td>Fragment 1, 2, 3 - AP to <i>Escherichia coli</i> MG1655 red.</Td></Tr></Table>
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<Br>
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<Br>
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PCR following fragment.<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 - Ap</Td><Td>Fragment 3 - Cm</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><Td>lacI-R(FRT)2<Br>mhpA-SaApR</Td><Td>p(SaApR)-lacIN<Br>mhpA-CmC6</Td>
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</Tr>
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<Tr>
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<Td>Templete</Td><Td>IF2<Br>FS2</Td><Td>Fragment 2-C<Br>Fragment 2-D</Td><Td>pSG76A2</Td><Td>lacI<Br>FRT-Cm2</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 18th</b><Br>
<b>September 18th</b><Br>
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PCR result.<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/f/fc/TMU-Tokyo_0917_1.jpg"><Br>
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1-3: Fragment 1, 4-6: Fragment 2,<Br>
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7-9: Fragment 3 - Ap, 1-3: Fragment 3 - Cm<Br>
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<Br>
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<Br>
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Mini prep following plasmid<Br>
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<Table><Tr><Td>pSB1C3-Km, pSB1A3</Td></Tr></Table>
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<Br>
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<Br>
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PCR following fragment.
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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>fimE</Td><Td>Fragment 1</Td><Td>Fragment 2</Td><Td>Fragment 3 - Cm</Td>
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</Tr>
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<Tr>
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<Td>Primer</Td><Td>pBAD-N9<Br>fimE-C-EcoRⅠ</Td><Td>lacA-IRR/IRL<Br>cynR-SaApR</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>pBAD<Br>fimE</Td><Td>IF2<Br>FS2</Td><Td>Fragment 2-C<Br>Fragment 2-D</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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<Br>
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<b>September 19th</b><Br>
<b>September 19th</b><Br>
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PCR result.<Br>
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<IMG Src="https://static.igem.org/mediawiki/2013/c/c4/TMU-Tokyo_0919.jpg"><Br>
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1: fimE, 2-3: Fragment 1, 4-5: Fragment 2,<Br>
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6-7: Fragment 3 - Cm<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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Revision as of 20:49, 27 September 2013



Experiment > 3rd week



September 15th
PCR result

1: Fragment 1, 2: Fragment 2, 3: Fragment 3 - Ap,
4: Fragment 3- Cm, 5: pBAD-fimE


Ligation fragment and vector.
Plasmid nameFragment 1-A+BFragment 2Fragment 3 - Ap
FragmentFragment 1-A
Fragment 1-B
Fragment 2Fragment 3 - Ap
VectorpSB1C3pSB1C3pSB1C3


Transformation following plasmid to Escherichia coli MG1655 red.
Fragment 1, 2, 3 - Ap


September 16th
Transformation result.



Mini prep following plasmid.
Fragment 1, 2, 3 - Ap, 3 - Cm, Km-Blunt
Result

1: Fragment 1, 2: Fragment 3 - Ap,
3-5: Fragment 3 - Cm, 6-8: Km-Blunt


PCR following fragment.
FragmentFragment 1Fragment 2Fragment 3 - ApFragment 3 - Cm
PrimerlacA-IRR/IRL
cynR-SaApR
araCHC
araA-TcC
lacI-R(FRT)2
mhpA-SaApR
p(SaApR)-lacIN
mhpA-CmC6
TempleteIF2
FS2
Fragment 2-C
Fragment 2-D
pSG76A2lacI
FRT-Cm2


September 17th
PCR result.

1-2: Fragment 1, 3-4: Fragment 2,
5-6: Fragment 3 - Cm, 8-10: Fragment 3 - Ap


Electro Poration
Fragment 1, 2, 3 - AP to Escherichia coli MG1655 red.


PCR following fragment.
FragmentFragment 1Fragment 2Fragment 3 - ApFragment 3 - Cm
PrimerlacA-IRR/IRL
cynR-SaApR
araCHC
araA-TcC
lacI-R(FRT)2
mhpA-SaApR
p(SaApR)-lacIN
mhpA-CmC6
TempleteIF2
FS2
Fragment 2-C
Fragment 2-D
pSG76A2lacI
FRT-Cm2


September 18th
PCR result.

1-3: Fragment 1, 4-6: Fragment 2,
7-9: Fragment 3 - Ap, 1-3: Fragment 3 - Cm


Mini prep following plasmid
pSB1C3-Km, pSB1A3


PCR following fragment.
FragmentfimEFragment 1Fragment 2Fragment 3 - Cm
PrimerpBAD-N9
fimE-C-EcoRⅠ
lacA-IRR/IRL
cynR-SaApR
araCHC
araA-TcC
R(SaApR)-lacIN
mhpA-CmC6
TempletepBAD
fimE
IF2
FS2
Fragment 2-C
Fragment 2-D
lacI
FRT-Cm


September 19th
PCR result.

1: fimE, 2-3: Fragment 1, 4-5: Fragment 2,
6-7: Fragment 3 - Cm


PCR following fragment. (2nd PCR)
September 20th
September 21th
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