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Team:TMU-Tokyo/Notebook experiment/2nd
From 2013.igem.org
Experiment > 2nd week
September 8th
PCR following fragment. (2nd PCR)
| Fragment | Fragment IF2 | Fragment FS2 |
| Primer | CmNR-Not FLP-R | CmC2 FLP-L |
| Templete | PtetR1 FLP | FLP Sm |
1: Fragment IF2, 2: Fragment FS2
PCR following fragment. (3rd PCR)
| Fragment | Fragment 1 |
| Primer | lacA-IRR/IRL cynR-SaApR |
| Templete | IF2 FS2 |
September 9th
PCR following fragment. (3rd PCR)
| Fragment | Fragment 1-A | Fragment 1-B | Fragment 1-C | Fragment 1-D |
| Primer | lacA-IRR/IRL cynR-SaApR | lacA-IRR/IRL cynR-SaApR | lacA-IRR/IRL cynR-SaApR | CmC2 CmNR-Not |
| Templete | IF2 FS | IF FS2 | IF2 FS2 | IF2 FS2 |
September 10th
PCR Result (Fragment 1-A, 1-B, 1-C, 1-D)
1: Fragment 1-A, 2: Fragment 1-B,
3: Fragment 1-C, 4: Fragment 1-D
Plasmid Extract attP (BBa_K617004)
Vector Remodeling with PCR. Insertion Recognition sites (NruⅠ).
| Fragment | C3 | attP1･2 | A3 |
| Primer | pSB1C3-NSP2 pSB1C3-NXE2 | attP-NruⅠR attP-NruⅠL | pSB1C3-NSP2 pSB1C3-NXE2 |
| Templete | attP | attP | IRR |
1: C3, 2: attP1･2, 3: A3
DNA Existract following fragment from gel.
| C3, A3, 1-A, 1-B, 1-3 |
September 11th
Digestion
| pSB1C3, pSB1A3 |
1: C3, 2:A3
Electro Pration
| Fragment 1, 2, 3 to Escherichia coli MG1655 red |
PCR following vector
| Fragment | pSB1A3 | pSB1C3 |
| Primer | pSB1C3-NSP2 pSB1C3-NXE2 | pSB1C3-NSP2 pSB1C3-NXE2 |
| Templete | attP IRR | attP IRR |
1-2: pSB1A3, 3-7: pSB1C3
September 11th
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