Team:Paris Liliane Bettencourt/Notebook/2010/07/05/
From 2010.igem.org
Raphaël and Théotime
Restriction digest - Xba I
- pSUlib - attC KanR
Final volume : 20 µL
15 µL distilled water
2 µL minipreped DNA
0,2 µL BSA 100x
2 µL buffer 10x
1 µL restriction enzyme
Protocol
1- Add in order listed above.
2- Vortex the mix 2 sec, spindown in micro-centrifuge and put in 37°C for 1h.
DNA Gel electrophoresis (agarose 1,5% w/v)
7,5g of agarose in 500mL of TBE buffer 1x
2 µL of EtBr was added in the gel
Protocol (2 wells)
- 10 µL of minipreped DNA + 2 µL of loading buffer 6x
- 10 µL of ladder 1Kb
Migration : 50V, 1h
Experiment failed, the migration went too far !! (no DNA visible in the well of minipreped DNA)
Antoine
Overnight culture 37°C
10mL LB + Kan
- pSUlib - attC KanR