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Team:UNICAMP-EMSE Brazil/Notebook/6 September 2011
From 2011.igem.org
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Gel 1: flhDC+HlyB+HlyD+TolC+T (C1); flhDC+HlyB+HlyD+TolC+T (C2); flhDC+HlyB+HlyD+TolC +T (C3); Sox S+ HlyB+HlyD+TolC (C1); RBS+ GFP+HlyA+ T (C2); RBS+ GFP+HlyA+ T (C3); flhDC+RBS+IL12 (C1).
06 September 2011
Checking digestion (E-P) of mini-prep constructs.
Objective:
- Checking digestion (E-P) of mini-preped constructs. It was used larger amount of DNA for the digestion.
Samples:
- Gel 1:
- flhDC+HlyB+HlyD+TolC+T (C1); flhDC+HlyB+HlyD+TolC+T (C2); flhDC+HlyB+HlyD+TolC +T (C3); Sox S+ HlyB+HlyD+TolC (C1); RBS+ GFP+HlyA+ T (C2); RBS+ GFP+HlyA+ T (C3); flhDC+RBS+IL12 (C1).
- Ladder:
- 100-10.000 bp (Fermentas)
- Gel Agarose concentration:
- 1,0%
- Results:
| Construction | Total size (gene+vector) | Linear vector size | Gene size | Result |
|---|---|---|---|---|
| flhDC+HlyB+HlyD+TolC+T (C1) | 8280 bp | 2800 bp | 5480 bp | OK |
| flhDC+HlyB+HlyD+TolC+T (C2) | 8280 bp | 2800 bp | 5480 bp | OK |
| flhDC+HlyB+HlyD+TolC+T (C3) | 8280 bp | 2800 bp | 5480 bp | Probably OK |
| SoxS+RBS+HlyB+HlyD+TolC + T (C1) | 8257 bp | 2800 bp | 5457 bp | NOT OK |
| RBS+GFP+HlyA+T (C2) | 4262 bp | 3205 bp | 1057 bp | OK |
| RBS+GFP+HlyA+T (C2) | 4262 bp | 3205 bp | 1057 bp | Smaller than expected |
| fLHDC+RBS+IL12 (C1) | 4584 bp | 2800 bp | 1784 bp | OK |
