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Thermo Scientific FastAP Thermosensitive Alkaline Phosphatase catalyzes the release of 5'- and 3'-phosphate groups from DNA, RNA, and nucleotides. This enzyme also removes phosphate groups from proteins.
FastAP is a novel alkaline phosphatase, which is active in all Thermo Scientific restriction enzyme buffers as well as in PCR buffers. It dephosphorylates all types of DNA ends (blunt, 5'- and 3'-overhangs) in 10 minutes at 37°C. The enzyme is inactivated in 5 minutes at 75°C (see Figure 1 in Supporting Data). Therefore, removal of alkaline phosphatase is not required prior to ligation.
Thermoinactivation curve of FastAP Thermosensitive Alkaline Phosphatase at 75°C.The DNA dephosphorylation reaction mixture (200 u/mL phosphatase) was incubated for 30 min at 37°C. Aliquots were heated at 75°C for time indicated on the graph. The remaining activity was measured by p-NPP assay.