|
|
| Line 13: |
Line 13: |
| | | | |
| | <b>September 22th</b><Br> | | <b>September 22th</b><Br> |
| - | PCR following fragment. (1st PCR)<Br>
| |
| - |
| |
| - | <div style="text-align:center;">
| |
| - | <Table cellpadding="5">
| |
| - | <Tr>
| |
| - | <Td>Fragment</Td><Td>PtetR1</Td><Td>PtetR2</Td><Td>FLP</Td><Td>Sm</Td><Td>araC-P</Td>
| |
| - | </Tr>
| |
| - | <Tr>
| |
| - | <Td>Primer</Td><Td>CmNR-stu<Br>FLP-IRR/IRL</Td><Td>CmCR-stu<Br>FLP-IRR/IRL</Td><Td>FLP-L<Br>FLP-R</Td><Td>FLP-SaApL<Br>CmC</Td><Td>acaC-2<Br>BAD-C3</Td>
| |
| - | </Tr>
| |
| - | <Tr>
| |
| - | <Td>Templete</Td><Td>IRR/IRL-GFP<Br>(BBa_K880002)</Td><Td>IRR/IRL-GFP<Br>(BBa_K880002)</Td><Td>pFTGT</Td><Td>m(C2-Ap)Sm</Td><Td>pBAD/Myc-HisA</Td>
| |
| - | </Tr>
| |
| - | </Table>
| |
| - | </div>
| |
| - |
| |
| - | <div style="text-align:center;">
| |
| - | <Table cellpadding="5">
| |
| - | <Tr>
| |
| - | <Td>Fragment</Td><Td>hbiF</Td><Td>Tc</Td><Td>lacI</Td><Td>FRT-Cm</Td><Td>FRT-Ap</Td>
| |
| - | </Tr>
| |
| - | <Tr>
| |
| - | <Td>Primer</Td><Td>pBAD-HbiF<Br>RTcN-hbiF</Td><Td>TcN<Br>TcC3</Td><Td>141-70<Br>FRT-lacIC</Td><Td>R6Kori-Xho-3R<Br>R(FRT)2</Td><Td>lacI-R(FRT)2<Br>mhpA-SaApR</Td>
| |
| - | </Tr>
| |
| - | <Tr>
| |
| - | <Td>Templete</Td><Td>hbiF<Br>(BBa_K880000)</Td><Td><i>Escherichia coli</i><Br>MG1655 recA⁻<Br>(cell suspention)</Td><Td><i>Escherichia coli</i><Br>MG1655 genome</Td><Td>pSG76C</Td><Td>pSG76A2</Td>
| |
| - | </Tr>
| |
| - | </Table>
| |
| - | </div>
| |
| - |
| |
| - |
| |
| - | Result<Br>
| |
| - | <IMG Src="https://static.igem.org/mediawiki/2013/f/f3/TMU-Tokyo_0902.jpg" width="350" hight="305"><Br>
| |
| - | 1: PterR1, 2: PtetR2, 3: FLP, 4: Sm, 5: araC-P,<Br>
| |
| - | 6: hbiF, 7: Tc, 8: LacI, 9: FRT-Cm, 10: FRT-Ap<Br>
| |
| - |
| |
| | | | |
| | | | |
| | + | <b>September 23th</b><Br> |
| | <Br> | | <Br> |
| | <Br> | | <Br> |
| | <b>September 24th</b><Br> | | <b>September 24th</b><Br> |
| - | Vector Remodeling with PCR. Insertion Recognition sites (Blunt).<Br>
| |
| - | <div style="text-align:center;">
| |
| - | <Table cellpadding="5">
| |
| - | <Tr>
| |
| - | <Td>Fragment</Td><Td>Km-Blunt</Td>
| |
| - | </Tr>
| |
| - | <Tr>
| |
| - | <Td>Primer</Td><Td>Km-Blunt-N<Br>Km-Blunt-C</Td>
| |
| - | </Tr>
| |
| - | <Tr>
| |
| - | <Td>Templete</Td><Td>pACYC177</Td>
| |
| - | </Tr>
| |
| - | </Table>
| |
| - | </div>
| |
| - |
| |
| - | Result<Br>
| |
| - | <IMG Src="https://static.igem.org/mediawiki/2013/1/19/TMU-Tokyo_0903.jpg" width="144" hight="250"><Br>
| |
| - | <Br>
| |
| - | <Br>
| |
| - | DNA Existract following fragment from gel.<Br>
| |
| - | <Table>
| |
| - | <Tr><Td>PtetR1, FLP, Sm, araC-P, hbiF, Tc, lacI, FRT-Cm, FRT-Ap, Km-Blunt</Td></Tr>
| |
| - | </Table>
| |
| - | <Br>
| |
| - | <Br>
| |
| - |
| |
| | | | |
| | <b>September 25th</b><Br> | | <b>September 25th</b><Br> |
| - | PCR following fragment. (2nd PCR)<Br>
| |
| | | | |
| - | <div style="text-align:center;">
| |
| - | <Table cellpadding="5">
| |
| - | <Tr>
| |
| - | <Td>Fragment</Td><Td>Fragment 1</Td><Td>Fragment 2</Td><Td>Fragment 3</Td>
| |
| - | </Tr>
| |
| - | <Tr>
| |
| - | <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>
| |
| - | </Tr>
| |
| - | <Tr>
| |
| - | <Td>Templete</Td><Td>PtetR<Br>FLP<Br>Sm</Td><Td>araC-P<Br>hbiF<Br>Tc</Td><Td>lacI<Br>FRT-Cm</Td>
| |
| - | </Tr>
| |
| - | </Table>
| |
| - | </div>
| |
| | | | |
| - | Result<Br>
| + | <b>September 26th</b><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<Br>
| + | |
| - | <Br>
| + | |
| - | <Br> | + | |
| | | | |
| - | <b>September 26th</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>
| |
| | | | |
| | <b>September 27th</b><Br> | | <b>September 27th</b><Br> |
"
