Dephosphorylation using FastDigest Restriction Enzymes and FastAP Alkaline Phosphatase

For years we have known that FastAP Alkaline Phosphatase is fully active in the FastDigest Buffers, both green and colorless versions. However, recent thorough examination of the enzymes revealed that dephosphorylation efficiency is highly dependent upon the nature of the restriction enzyme used in digestion.

With some enzymes, restriction digestion and dephosphorylation reactions can be performed simultaneously. Others may require heat inactivation or column purification following restriction enzyme digestion and prior to the addition of FastAP Alkaline Phosphatase.

As a result, all FastDigest Restriction Enzymes have been tested to determine the optimal protocol for complete dephosphorylation when using FastAP Alkaline Phosphatase. The recommendations are outlined in the table below.