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Team:UPO-Sevilla/Notebook/08 13
From 2010.igem.org
August, 13th
Assembly Team
There were kept inocula of verified devices to save them at -80ºC. We made plasmid purifications by using those inocula and we analyzed them again by EcoRI and PstI cutting and agarose gel electrophoresis. Sadly, not all the analysis showed positive results. Then we made other analytic digestions to check those devices and the day before devices.
| PARTS | RESTRICTION ENZYMES | PART SIZES | FRAGMENT SIZES | RESULTS |
|---|---|---|---|---|
| UPO2 | SspI + PstI | 15 | 1892, 271 | ✔ |
| 1+2 | SspI + PstI | 35+15 | 1892, 315 | ✔ |
| pSB1A3 (control) | SspI + PstI | 2157 | 1912, 340+1200 | ✔ |
| UPO12 (pMA) | PstI + NruI | 86 | it would be just one cut | ✔ |
| 12+2 | PstI + NruI | 86+15 | 1995, 589 | ✕ |
| pSB1T3 (control) | PstI + NruI | 2416 | 1995, 488+1200 | ✔ |
| UPO19 (1AK3) | EcoRI + PvuII | 1892 | 1077, 815+3189 | ✔ |
| 1+19 (1T3) | EcoRI + PvuII | 35+1892 | 1112, 815+2463 | ? |
| UPO19 (1AK3) | PvuII + PstI | 1892 | 815, 1077+3189 | ✔ |
| 1+19 (1T3) | PvuII + PstI | 35+1892 | 815, 1112+2463 | ✔ |
| 11+2+16+3 (3C5) | EcoRI + BglII | 192+15+1518+129 | 279, 1239+V | ✕ |
| 12+2+16+3 (3C5) | EcoRI + BglII | 86+15+1518+129 | 173, 1239+V | ✕ |
| 16+3 (1C3) | EcoRI + BglII | 1518+129 | 72, 1239+V | ? |
| 2+4+2+6 | pending | pending | pending | ✕ |
| 1+2+4 | EcoRI + SphI | 35+15+2316 | 716, 1650+V | ✕ |
| 2+4 | EcoRI + SphI | 35+15+2316 | 661, 1650+V | ✕ |
| 1+2+7+2 | EcoRI + SspI | 35+15+2292+15 | 161, 2181+V | ✕ |
| 7+2 | EcoRI + SspI | 2292+15 | 111 ,2181+V | ✔ |
| 11+19 | EcoRI + EcoRV | 192 + 1892 | 280, 1804+V | ✕ |
| 19 | 1892 | 88, 1804+V | ✔ | |
| 12+2+13+3 | EcoRI + NcoI | 86+15+720+129 | 216, 554+V | ✕ |
| 13+3 | EcoRI + NcoI | 720+129 | 166, 554+V | ✕ |
| UPO28 | SphI + PstI | 1472 | 1121, 351+V | ✔ |
| UPO28 | SphI + EcoRI | 1472 | 351, 1121+V | ✕ |
| 2+18 | EcoRI +SphI | pending | pending | pending |
| 18 | EcoRI +SphI | pending | pending | pending |
There we had three kinds of results:
- Confirmed devices: 1+2, 7+2, 13+3.
- Not assembled devices: 12+2, 2+4, 28, 11+19, 1+2+4, 11+2+16+3, 12+2+16+3, 1+2+7+2, 12+2+13+3.
- Need another analysis: 1+19, 16+3.
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