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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)

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Antoine

Overnight culture 37°C

10mL LB + Kan

• pSUlib - attC KanR