Dephosphorylation using CIP in Restriction Enzyme Reaction ( M0290 )
Digest 1–5 μg of plasmid DNA in a 20 μl reaction as follows : DNA ≥ 1 μl ; Restriction Enzyme Buffer ( 10X ) 2 μl ; Restriction Endonuclease 1 μl ; H2O , purified to 20 μl .
Incubate at 37°C for 60 minutes or follow manufacturer’s recommendations .
Add 1 unit of CIP for every 1 pmol of DNA ends ( about 1 μg of a 3 kb plasmid ) .
Incubate at 37°C for 30–60 minutes .
Purify DNA by gel purification , spin - column or phenol extraction .
Proceed with ligation .
