Shrimp alkaline phosphatase, recombinant (rSAP) is a heat-labile hydrolase enzyme produced in Pichia pastoris that removes phosphate groups non specifically from 5’ ends of nucleic acid phosphomonoesters and proteins. This activity is most commonly utilised in molecular cloning to prevent self-ligation of linearised plasmid DNA and in 5’ end-labelling to facilitate the replacement of unlabelled phosphates with labeled phosphate groups. rSAP also prepares PCR products for DNA sequencing or SNP analysis, by dephosphorylating unincorporated dNTPs that would otherwise interfere with enzymatic reactions.
- Removes 5’-phosphates from DNA, RNA, dNTPs, and proteins
- Improves cloning efficiency by preventing vector recircularisation
- 100% heat-inactivated at 65 °C, no vector purification necessary
- Removes unincorporated dNTPs in PCR products prior to DNA sequencing or SNP analysis
- Prepares templates for 5’ end labelling
- Dephosphorylates proteins
- Works in many different buffers without supplemental factors
rSAP may be directly added to restriction enzyme digests and is conveniently 100% inactivated by heating at 65 °C. This eliminates the need for vector purification, a necessary step when using alkaline phosphatases isolated from other sources, such as E. coli and calf intestine. rSAP works well in common buffers and does not require supplemental zinc or other additives.