Team:UPO-Sevilla/Notebook/08 13

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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

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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