Dephosphorylation using AnP ( M0289 )
Add 1 / 10 volume of 10X Antarctic Phosphatase Reaction Buffer to 1 - 5 µg of DNA cut with any restriction endonuclease in any buffer .
Add 1 µl of Antarctic Phosphatase ( 5 units ) and mix .
Incubate for 15 minutes at 37°C for 5´ extensions or blunt - ends , 60 minutes for 3´ extensions .
Heat inactivate ( or as required to inactivate the restriction enzyme ) for 5 minutes at 70°C .
Proceed with ligation .
