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Pyrophosphatase, Inorganic (from yeast)*
* Use of this enzyme in certain applications may be covered by patents and may require a license

#EF0221 Supplied with: Storage (Dilution) Buffer	10 u (0.1 u/µl) 1 ml
Related Documents (in pdf, ~40-60 KB): Certificate of Analysis: #EF0221 MSDS (English) MSDS (English-USA) MSDS (German)

Description
The Pyrophosphatase, Inorganic (PPase) catalyzes the hydrolysis of inorganic pyrophosphate to two orthophosphates. The enzyme requires a divalent metal cation, with Mg²⁺ conferring the highest activity (1).

Source
E.coli cells with a cloned ppa gene of Sacharomyces cerevisiae.

Molecular Weight
This enzyme is homodimer. It consists of two identical subunits of 32 kDa.

Applications

• High yield synthesis of RNA by in vitro transcription (2).
• DNA polymerization reactions: preventing accumulation of pyrophosphate (3, 4).
• Removal of contaminant PPi in reagents used for SNP genotyping by methods based on the detection of pyrophosphate (5).

Quality Control
The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests.

Concentration
0.1 u/µl

Definition of Activity Unit
One unit of the enzyme hydrolyzes 1 µmol of inorganic pyrophosphate in 1 min at 25°C.
Enzyme activity is assayed in the following mixture: 100 mM Tris-HCl (pH 7.2), 2 mM MgCl₂ and 2 mM inorganic pyrophosphate (PPi).

Storage (Dilution) Buffer
The enzyme is supplied in: 10 mM Tris-HCl (pH 7.5), 0.1 mM EDTA and 50% (v/v) glycerol.

Inhibition and Inactivation
Inactivated by phenol/chloroform extraction.

Note

• Recommended concentration of PPase in RNA synthesis is 1-5 u/ml of transcription reaction mixture.
• The enzyme can be diluted with supplied storage (dilution) buffer.

Related Products

phi29 DNA Polymerase Klenow Fragment Klenow Fragment, exo- RNA Polymerases M-MuLV Reverse Transcriptase RevertAid™ M-MuLV Reverse Transcriptase RevertAid™ H Minus M-MuLV Reverse Transcriptase Water, nuclease-free

Reference

1. Cooperman, B.S., The mechanism of action of yeast inorganic pyrophosphatase, Meth. Enzymol., 87, 526-548, 1982.
2. Cunningham, P.R. and Ofengand, J., Use of inorganic pyrophostase to improve the yield of in vitro transcription reactions catalyzed by T7 RNA polymerase, Biotechniques, 9, 713-714, 1990.
3. Tabor, S., Richardson, C.C., DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Effect of pyrophosphorolysis and metal ions, J. Biol. Chem., 265, 8322-8328, 1990.
4. Dean, B.F., et al., Rapid amplification of plasmid and phage DNA using phi29 DNA polymerase and multiply-primed Rolling Circle amplification, Genome Res., 11, 1095-1099, 2001.
5. Zhou, G.H., et al., Quantitative detection of single nucleotide polymorphisms for a pooled sample by a bioluminometric assay coupled with modified primer extension reactions (BAMPER), Nucleic Acids Res., 29, E93, 2001.

Updated vasario 05, 2008 14:35