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Team:TMU-Tokyo/Notebook experiment/1st
From 2013.igem.org
(Difference between revisions)
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</Table> | </Table> | ||
Result<Br> | Result<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/d/dd/TMU-Tokyo_09051.jpg"><Br> | ||
| + | 1: PtetR1, 2: FLP, 3: Sm, 4: araC-P, 5: hbiF,<Br> | ||
| + | 6: Tc, 7: lacI, 8: FRT-Cm, 9: FRT-Ap, 10: Km-Blunt<Br> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | |||
| + | |||
| + | |||
| + | <b>September 6th</b><Br> | ||
| + | PCR following fragment. (2nd PCR)<Br> | ||
| + | <div style="text-align:center;"> | ||
| + | <Table cellpadding="5"> | ||
| + | <Tr> | ||
| + | <Td>Fragment</Td><Td>Fragment 1-A</Td><Td>Fragment 1-B</Td><Td>Fragment 2-A</Td><Td>Fragment 2-B</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <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> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Templete</Td><Td>PtetR<Br>FLP</Td><Td>FLP<Br>Sm</Td><Td>Sm<Br>FLP</Td><Td>Sm<Br>FLP</Td> | ||
| + | </Tr> | ||
| + | </Table> | ||
| + | </div> | ||
| + | Result<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/a/a0/TMU-Tokyo_0906.jpg"><Br> | ||
| + | 1: Fragment 1-A, 2: Fragment 2-B<Br> | ||
| + | (1-B and 2-A was failed in PCR)<Br> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | PCR following fragment. (2nd PCR)<Br> | ||
| + | <div style="text-align:center;"> | ||
| + | <Table cellpadding="5"> | ||
| + | <Tr> | ||
| + | <Td>Fragment</Td><Td>Fragment IF</Td><Td>Fragment FS</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Primer</Td><Td>lacA-IRR/IRL<Br>FLP-R</Td><Td>FLP-L<Br>cynR-SaApR</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Templete</Td><Td>PtetR<Br>FLP</Td><Td>FLP<Br>Sm</Td> | ||
| + | </Tr> | ||
| + | </Table> | ||
| + | </div> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | |||
| + | <b>September 7th</b><Br> | ||
| + | PCR Result (Fragment IF, FS)<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/8/89/TMU-Tokyo_0907.jpg"><Br> | ||
| + | 1: Fragment IF, 2: Fragment FS<Br> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | PCR following fragment. (3rd PCR)<Br> | ||
| + | <div style="text-align:center;"> | ||
| + | <Table cellpadding="5"> | ||
| + | <Tr> | ||
| + | <Td>Fragment</Td><Td>Fragment 1</Td><Td>Fragment 2</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Primer</Td><Td>lacA-IRR/IRL<Br>cynR-SaApR</Td><Td>araCHC<Br>araA-TcC</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Templete</Td><Td>Fragment IF<Br>Fragment FS</Td><Td>Fragment 2-B<Br>Fragment 2-C</Td> | ||
| + | </Tr> | ||
| + | </Table> | ||
| + | </div> | ||
| + | Result<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/5/52/TMU-Tokyo_09071.jpg"><Br> | ||
| + | 1: Fragment 1, 2: Fragment 2<Br> | ||
<Br> | <Br> | ||
<Br> | <Br> | ||
<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 17:39, 27 September 2013
Experiment > Parts Synthesis > 1st week
September 2nd
PCR following fragment. (1st PCR)
| Fragment | PtetR1 | PtetR2 | FLP | Sm | araC-P |
| Primer | CmNR-stu FLP-IRR/IRL | CmCR-stu FLP-IRR/IRL | FLP-L FLP-R | FLP-SaApL CmC | acaC-2 BAD-C3 |
| Templete | IRR/IRL-GFP (BBa_K880002) | IRR/IRL-GFP (BBa_K880002) | pFTGT | m(C2-Ap)Sm | pBAD/Myc-HisA |
| Fragment | hbiF | Tc | lacI | FRT-Cm | FRT-Ap |
| Primer | pBAD-HbiF RTcN-hbiF | TcN TcC3 | 141-70 FRT-lacIC | R6Kori-Xho-3R R(FRT)2 | lacI-R(FRT)2 mhpA-SaApR |
| Templete | hbiF (BBa_K880000) | Escherichia coli MG1655 recA⁻ (cell suspention) | Escherichia coli MG1655 genome | pSG76C | pSG76A2 |
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).
| Fragment | Km-Blunt |
| Primer | Km-Blunt-N Km-Blunt-C |
| Templete | pACYC177 |
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)
| Fragment | Fragment 1 | Fragment 2 | Fragment 3 |
| Primer | lacA-IRR/IRL cynR-SaApL | araCHC araA-TcC | R(SaApR)-lacIN mhpA-CmC6 |
| Templete | PtetR FLP Sm | araC-P hbiF Tc | lacI FRT-Cm |
1: Fragment 1, 2: Fragment 2, 3: Fragment 3
September 5th
PCR following fragment. (2nd PCR)
| Fragment | Fragment 1-A | Fragment 1-B | Fragment 2-C | Fragment 2-D |
| Primer | CmNR-Stu FLP-R | FLP-L CmC | araC2 RTcN-hbiF | pBAD-hbiF TcC3 |
| Templete | PtetR FLP | FLP Sm | araC-P hbiF | hbiF Tc |
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 |
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)
| Fragment | Fragment 1-A | Fragment 1-B | Fragment 2-A | Fragment 2-B |
| Primer | CmNR-Stu FLP-R | FLP-L CmC | FLP-L CmC | FLP-L cynR-SaApL |
| Templete | PtetR FLP | FLP Sm | Sm FLP | Sm FLP |
1: Fragment 1-A, 2: Fragment 2-B
(1-B and 2-A was failed in PCR)
PCR following fragment. (2nd PCR)
| Fragment | Fragment IF | Fragment FS |
| Primer | lacA-IRR/IRL FLP-R | FLP-L cynR-SaApR |
| Templete | PtetR FLP | FLP Sm |
September 7th
PCR Result (Fragment IF, FS)
1: Fragment IF, 2: Fragment FS
PCR following fragment. (3rd PCR)
| Fragment | Fragment 1 | Fragment 2 |
| Primer | lacA-IRR/IRL cynR-SaApR | araCHC araA-TcC |
| Templete | Fragment IF Fragment FS | Fragment 2-B Fragment 2-C |
1: Fragment 1, 2: Fragment 2
>>Go to next week
