IGEM:Harvard/2007/Protocols/Vector Dephosphorylation Protocol
- Add 1/10 volume of 10X Antarctic Phosphatase Reaction Buffer to 1 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 65°C.
- Proceed with ligation.