Thermo Scientific FastAP thermosensitive alkaline phosphatase catalyzes 5'- and 3' release from DNA, RNA and nucleotides -Phosphate group. The enzyme also removes phosphate groups from proteins.

FastAP is a novel alkaline phosphatase active in all Thermo Scientific restriction enzyme buffers and PCR buffers. It dephosphorylates all types of DNA ends (blunt ends, 5'- and 3'-overhangs) in 10 minutes at 37°C. The enzyme is inactivated within 5 minutes at 75°C (SeeFigure 1 in the Supporting Data). Therefore, alkaline phosphatase does not need to be removed prior to ligation.

Product Advantages

• Recombinase
• Rapid dephosphorylation—37°C 10 minutes
• Fast and complete inactivation—5 minutes at 75°C
• Simultaneous digestion and dephosphorylation of vector DNA
• 100% active in restriction enzymes and PCR buffers
• PCR purification binds Exo I
• Protein dephosphorylation

One solution for all types of DNA ends:
•5'-overhang
• 3'-overhang
• Flat Terminal
• Single Nucleotide

Applications

• Dephosphorylation of cloning vector DNA to prevent recircularization during ligation
• Simultaneous digestion and dephosphorylation of vector DNA
• PCR product purification: nucleotide degradation before PCR product sequencing
• Dephosphorylation of nucleic acid'-ends before labeling by T4 polynucleotide kinase 5
• Other applications where dephosphorylation of DNA and RNA substrates must be performed
• Protein dephosphorylation Phosphorylation

Note

• Binding of FastAP thermosensitive alkaline phosphatase to DNA may cause band shifts in agarose gels. To avoid this, use 6X DNA Loading Dye & SDS Solution at 65°C Incubate the samples for 10 minutes, cool on ice and perform electrophoresis.
•FastAP thermosensitive alkaline phosphatase is active in all restriction enzyme buffers and can be added directly to digested DNA. No heat-inactivated restriction enzymes are required before dephosphorylation.