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Team:TMU-Tokyo/Notebook experiment/1st
From 2013.igem.org
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<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> | ||
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<IMG Src="https://static.igem.org/mediawiki/2013/1/19/TMU-Tokyo_0903.jpg" width="144" hight="250"><Br> | <IMG Src="https://static.igem.org/mediawiki/2013/1/19/TMU-Tokyo_0903.jpg" width="144" hight="250"><Br> | ||
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<IMG Src="https://static.igem.org/mediawiki/2013/d/d1/TMU-Tokyo_0904.jpg"><Br> | <IMG Src="https://static.igem.org/mediawiki/2013/d/d1/TMU-Tokyo_0904.jpg"><Br> | ||
| - | 1: Fragment 1, 2: Fragment 2, 3: Fragment 3 | + | 1: Fragment 1, 2: Fragment 2, 3: Fragment 3<Br> |
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| + | <b>September 5th</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-C</Td><Td>Fragment 2-D</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <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> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Templete</Td><Td>PtetR<Br>FLP</Td><Td>FLP<Br>Sm</Td><Td>araC-P<Br>hbiF</Td><Td>hbiF<Br>Tc</Td> | ||
| + | </Tr> | ||
| + | </Table> | ||
| + | </div> | ||
| + | Result<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/9/9c/TMU-Tokyo_0905.jpg"><Br> | ||
| + | 1: Fragment 1-A, 2: Fragment 1-B,<Br> | ||
| + | 3: Fragment 2-C, 4: Fragment 2-D<Br> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | Check following gel extract product.<Br> | ||
| + | <Table> | ||
| + | <Tr><Td>PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt</Td></Tr> | ||
| + | </Table> | ||
| + | Result<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 16:59, 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 |
>>Go to next week
