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.
