Team:TU Darmstadt/protocols/Restriction digest


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Restriction digest

Short explanation
In order to insert DNA fragments into plasmids via ligation it is necessary to digest both components with restriction enzymes.

DNA Digestion (50 µL batch)
Add up the following:
10x NEBuffer 5 µL
DNA 1 µg
Restriction Enzyme (10 u) 1 µL
100x BSA 0.2 µL
Add dH2O for total reaction volume of 50 µL

Incubate for 1 h at an temperatur of 37°C
inactivate enzymes by incubating at 65°C for 20 min