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  • Protocol for Dephosphorylation of 5´-ends of DNA using Antarctic Phosphatase (M0289)

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    1. Prepare a 20 μl reaction as follows:

      DNA 1 pmol of DNA ends*
      Antarctic Phosphatase Rxn Buffer (10X) 2 μl
      Antarctic Phosphatase 5 units
      H2O, purified to 20 μl**

    2. Incubate at 37°C for 30 minutes.

    3. Stop reaction by heat-inactivation at 80°C for 2 minutes.

    * Note: 1 pmol of DNA ends is about 1 μg of a 3 kb plasmid.
    ** Scale larger reaction volumes proportionally.