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Team:TMU-Tokyo/Notebook experiment/3rd
From 2013.igem.org
(Difference between revisions)
| Line 81: | Line 81: | ||
<b>September 17th</b><Br> | <b>September 17th</b><Br> | ||
| + | PCR result.<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/7/7c/TMU-Tokyo_0917.jpg"><Br> | ||
| + | 1-2: Fragment 1, 3-4: Fragment 2,<Br> | ||
| + | 5-6: Fragment 3 - Cm, 8-10: Fragment 3 - Ap<Br> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | Electro Poration<Br> | ||
| + | <Table><Tr><Td>Fragment 1, 2, 3 - AP to <i>Escherichia coli</i> MG1655 red.</Td></Tr></Table> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | PCR following fragment.<Br> | ||
| + | <div style="text-align:center;"> | ||
| + | <Table cellpadding="5"> | ||
| + | <Tr> | ||
| + | <Td>Fragment</Td><Td>Fragment 1</Td><Td>Fragment 2</Td><Td>Fragment 3 - Ap</Td><Td>Fragment 3 - Cm</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <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> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <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> | ||
| + | </Tr> | ||
| + | </Table> | ||
| + | </div> | ||
| + | <Br> | ||
| + | <Br> | ||
| Line 87: | Line 113: | ||
<b>September 18th</b><Br> | <b>September 18th</b><Br> | ||
| + | PCR result.<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/f/fc/TMU-Tokyo_0917_1.jpg"><Br> | ||
| + | 1-3: Fragment 1, 4-6: Fragment 2,<Br> | ||
| + | 7-9: Fragment 3 - Ap, 1-3: Fragment 3 - Cm<Br> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | Mini prep following plasmid<Br> | ||
| + | <Table><Tr><Td>pSB1C3-Km, pSB1A3</Td></Tr></Table> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | |||
| + | PCR following fragment. | ||
| + | <div style="text-align:center;"> | ||
| + | <Table cellpadding="5"> | ||
| + | <Tr> | ||
| + | <Td>Fragment</Td><Td>fimE</Td><Td>Fragment 1</Td><Td>Fragment 2</Td><Td>Fragment 3 - Cm</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <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> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <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> | ||
| + | </Tr> | ||
| + | </Table> | ||
| + | </div> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | |||
| + | |||
<b>September 19th</b><Br> | <b>September 19th</b><Br> | ||
| + | PCR result.<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/c/c4/TMU-Tokyo_0919.jpg"><Br> | ||
| + | 1: fimE, 2-3: Fragment 1, 4-5: Fragment 2,<Br> | ||
| + | 6-7: Fragment 3 - Cm<Br> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | PCR following fragment. (2nd PCR)<Br> | ||
| + | |||
| + | |||
| + | |||
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 name | Fragment 1-A+B | Fragment 2 | Fragment 3 - Ap |
| Fragment | Fragment 1-A Fragment 1-B | Fragment 2 | Fragment 3 - Ap |
| Vector | pSB1C3 | pSB1C3 | pSB1C3 |
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 |
1: Fragment 1, 2: Fragment 3 - Ap,
3-5: Fragment 3 - Cm, 6-8: Km-Blunt
PCR following fragment.
| Fragment | Fragment 1 | Fragment 2 | Fragment 3 - Ap | Fragment 3 - Cm |
| Primer | lacA-IRR/IRL cynR-SaApR | araCHC araA-TcC | lacI-R(FRT)2 mhpA-SaApR | p(SaApR)-lacIN mhpA-CmC6 |
| Templete | IF2 FS2 | Fragment 2-C Fragment 2-D | pSG76A2 | lacI 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.
| Fragment | Fragment 1 | Fragment 2 | Fragment 3 - Ap | Fragment 3 - Cm |
| Primer | lacA-IRR/IRL cynR-SaApR | araCHC araA-TcC | lacI-R(FRT)2 mhpA-SaApR | p(SaApR)-lacIN mhpA-CmC6 |
| Templete | IF2 FS2 | Fragment 2-C Fragment 2-D | pSG76A2 | lacI 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.
| Fragment | fimE | Fragment 1 | Fragment 2 | Fragment 3 - Cm |
| Primer | pBAD-N9 fimE-C-EcoRⅠ | lacA-IRR/IRL cynR-SaApR | araCHC araA-TcC | R(SaApR)-lacIN mhpA-CmC6 |
| Templete | pBAD 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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