Uses
- Use rAPid Alkaline Phosphatase in numerous applications, including: Removal of 5′-phosphoryl groups from nucleic acids
- Preparation of templates for 5′-end labeling
- Clean-up of PCR products by removal of dNTPs
- Dephosphorylation of proteins
Note: rAPid Alkaline Phosphatase is very rapidly inactivated by heating at +75 °C for two minutes. No additional purification steps are required after restriction and dephosphorylation. The dephosphorylated DNA can directly be used in a ligation reaction.
Note: rAPid Alkaline Phosphatase is also included in the Rapid DNA Dephos & Ligation Kit, which enables fast and efficient dephosphorylation and ligation of sticky- or blunt-end DNA fragments. With this convenient kit, dephosphorylation of blunt and sticky 5′-overhanging ends is performed in ten minutes at +37 °C and ligation is completed in just five minutes at +15–25 °C.
General Description
rAPid Alkaline Phosphatase is an alkaline phosphatase isolated from bovine intestine and supplied as a recombinant enzyme of 56 kD, expressed in the yeast, Pichia Pastoris. The recombinant form ensures consistency and safety. rAPid Alkaline Phosphatase catalyzes the dephosphorylation of 5′ phosphates from DNA and RNA, nucleotides, and proteins. Alkaline phosphatase treatment prevents self-ligation of DNA fragments by removing the 5′-phosphoryl termini required by ligases; this feature is of major importance in cloning strategies to decrease vector background. Unlike calf intestinal phosphatase, rAPid Alkaline Phosphatase is rapidly, completely, and irreversibly inactivated by heat treatment for two minutes at +75 °C. It is therefore, an excellent alternative to Shrimp Alkaline Phosphatase. In addition, the enzyme is active in restriction enzyme buffers. Therefore, restriction enzyme digestion, dephosphorylation, enzyme inactivation, ligation, or 5′-end labeling can be performed without purification steps.
