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Team:TMU-Tokyo/Notebook experiment/4th
From 2013.igem.org
(Difference between revisions)
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<IMG Src="https://static.igem.org/mediawiki/2013/5/56/TMU-Tokyo_0924_2.jpg"><Br> | <IMG Src="https://static.igem.org/mediawiki/2013/5/56/TMU-Tokyo_0924_2.jpg"><Br> | ||
<IMG Src="https://static.igem.org/mediawiki/2013/b/b4/TMU-Tokyo_0924_3.jpg" width="250" Height="125"><Br> | <IMG Src="https://static.igem.org/mediawiki/2013/b/b4/TMU-Tokyo_0924_3.jpg" width="250" Height="125"><Br> | ||
| - | <Br> | + | Get Fragment 1-2<Br> |
<Br> | <Br> | ||
Electro Poration<Br> | Electro Poration<Br> | ||
| Line 167: | Line 167: | ||
<b>September 25th</b><Br> | <b>September 25th</b><Br> | ||
| - | + | Check PCR.<Br> | |
| + | <div style="text-align:center;"> | ||
| + | <Table cellpadding="5"> | ||
| + | <Tr> | ||
| + | <Td>Fragment</Td><Td>Fragment 3 - ApCm<Br>(control)</Td><Td>Fragment 3 - ApCm</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Primer</Td><Td>141-49<Br>141-1</Td><Td>CmNR-Not<Br>141-1</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Templete</Td><Td>(Cell suspention)</Td><Td>(Cell suspention)</Td> | ||
| + | </Tr> | ||
| + | </Table> | ||
| + | </div> | ||
| + | Result.<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/8/8b/TMU-Tokyo_0925.jpg"><Br> | ||
| + | 1-7: 141-49 and 141-1, 8: control(141-49 and 141-8),<Br> | ||
| + | 9-15: 141-1 and CmNR-Not, 16: control(141-1 and CmNR-Not)<Br> | ||
| + | <Br> | ||
| + | <Br> | ||
<b>September 26th</b><Br> | <b>September 26th</b><Br> | ||
| - | + | P1 transduction<Br> | |
| + | <Table><Tr><Td>Fragment 3 to Fragment 1-2</Td></Tr></Table> | ||
| + | Get Fragment 1-2-3.<Br> | ||
| + | <Br> | ||
| + | <Br> | ||
| + | Check PCR<Br> | ||
| + | <div style="text-align:center;"> | ||
| + | <Table cellpadding="5"> | ||
| + | <Tr> | ||
| + | <Td>Fragment</Td><Td>Fragment 1-2-3</Td><Td>Fragment 1-2<Br>(control)</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Primer</Td><Td>CmNR-Not<Br>141-1</Td><Td>141-49<Br>141-1</Td> | ||
| + | </Tr> | ||
| + | <Tr> | ||
| + | <Td>Templete</Td><Td>(Cell suspention)</Td><Td>(Cell suspention)</Td> | ||
| + | </Tr> | ||
| + | </Table> | ||
| + | </div> | ||
| + | Result.<Br> | ||
| + | <IMG Src="https://static.igem.org/mediawiki/2013/8/8a/TMU-Tokyo_0926.jpg"><Br> | ||
| + | 1-7: Fragment 1-2-3, 8: cotrol, 9-10: Fragment 1,<Br> | ||
| + | 11-13: Fragment 2, 14-15: Fragment 3 - Ap, 16-17: Fragment 3 - Cm<Br> | ||
| + | 18-19: Fragment 3 - AmCm<Br> | ||
| + | <Br> | ||
<b>September 27th</b><Br> | <b>September 27th</b><Br> | ||
Revision as of 23:51, 27 September 2013
Experiment > 4th week
September 22th
Mini prep following plasmid.
| fimE-pBAD(in pSB1A3), pSB1C3-Km, pSB1A3-Km |
1: pSB1C3-Km, 2-4: pSB1A3-Km, 5-6: fimE-pBAD
Check PCR.
| Fragment | (control) | Fragment 3 - ApCm |
| Primer | 141-49 SaApL | 141-49 SaApL |
| Templete | Escherichia coli MG1655 red (Cell suspention) | (Cell suspention) |
1-9: Fragment 3 - ApCm
Check PCR.
| Fragment | (control) | Fragment 3 - ApCm | Fragment 3 - ApCm |
| Primer | 2 sets of the others | 141-1 141-20 | 141-1 141-74 |
| Templete | Escherichia coli MG1655 red (Cell suspention) | (Cell suspention) |
1-7: 141-1 and 141-20, 8: control(141-1 and 141-20),
9-15: 141-1 and 141-74, 16: control(141-1 and 141-74),
Digestion following plasmid.
| pSB1C3-Km-Blunt |
P1 Preparation following fragment.
| Fragment 1, 2 |
September 23th
PCR following fragment.
| Fragment | Fragment 3 - ApCm |
| Primer | mhpA-CmC6 SaApL |
| Templete | Fragment 3 - Cm Ap(SaApR-SaApL) |
1: Fragment 3 - Cm, 2: Fragment 3 - ApCm,
3: pSB1C3-Km-Blunt
Check PCR.
| Fragment | Fragment 3 - Ap (control) | Fragment 3 - ApCm |
| Primer | CmNR-Not SaApL | CmNR-Not SaApL |
| Templete | (Cell suspention) | (Cell suspention) |
1-31: Fragment 3 - ApCm
Electro Pration
| Fragment 3 - Cm to Escherichia coli MG1655 red. |
P1 Preparation
| Fragment 3 - ApCm |
September 24th
P1 transdunction
| Fragment 1, 2 |
| Fragment | Fragment 3 - Ap (control) | Fragment 3 - ApCm |
| Primer | CmNR-Not 141-1 | CmNR-Not 141-1 |
| Templete | (Cell suspention) | (Cell suspention) |
1-16: Fragment 3 - ApCm
P1 transduction
| Fragment 2 to Fragment 1 |
| Fragment 1 to Fragment 2 |
Get Fragment 1-2
Electro Poration
| Fragment 1, 2, 3 - Ap, 3 - Cm, 3 - ApCm (in pSBC3-Km) to Escherichia coli DH1 |
September 25th
Check PCR.
| Fragment | Fragment 3 - ApCm (control) | Fragment 3 - ApCm |
| Primer | 141-49 141-1 | CmNR-Not 141-1 |
| Templete | (Cell suspention) | (Cell suspention) |
1-7: 141-49 and 141-1, 8: control(141-49 and 141-8),
9-15: 141-1 and CmNR-Not, 16: control(141-1 and CmNR-Not)
September 26th
P1 transduction
| Fragment 3 to Fragment 1-2 |
Check PCR
| Fragment | Fragment 1-2-3 | Fragment 1-2 (control) |
| Primer | CmNR-Not 141-1 | 141-49 141-1 |
| Templete | (Cell suspention) | (Cell suspention) |
1-7: Fragment 1-2-3, 8: cotrol, 9-10: Fragment 1,
11-13: Fragment 2, 14-15: Fragment 3 - Ap, 16-17: Fragment 3 - Cm
18-19: Fragment 3 - AmCm
September 27th
September 28th
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