Team:Lyon-INSA-ENS/Realisation/Protocols/Fermentas

Fermentas digestion

This protocol aims at cutting plasmids on specific restriction sites using restriction enzymes.

Procedure

1. In an eppendorf tube, add the following solutions in any order :
-250 ng DNA ( if the concentration is unknown, use 5µL )
-2.5µL (1X) or 5µL (2X) 10X tango buffer ( this depends on the enzymes used )
-water to get a volume of 25mL after addition of the enzymes

2. Add 1µL of each one of the desired enzymes.

3. Incubate at 37°C for 1h30

4. Incubate at 70°C for 10 mn to inactivate the restriction enzymes.