Thermostable Inorganic PyrophosphataseProduct information
- Retains 100% activity after incubation at 100°C for 4 hours
P2O7-4 + H2O -> 2HPO4-2.
Product SourceAn E. coli strain carrying a plasmid encoding a pyrophosphatase from the extreme thermophile Thermococcus litoralis.
Properties and Usage
Unit DefinitionOne unit is the amount of enzyme that will generate 1 μmol of phosphate per minute from inorganic pyrophosphate under standard reaction conditions (a 10 minute reaction at 75°C in 50 mM Tricine [pH 8.5] , 1 mM MgCl2, 0.32 mM PPi, reaction volume of 0.5 ml).
20 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
pH 8.0 @ 25°C
Quality Control AssaysThe following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
- dNTPase Activity:
The Pyrophosphatase is tested in a reaction containing a mix of dNTP's. After incubation the amount of inorganic phosphatase released is determined by comparison to a standard curve of known phosphate concentrations.
- Endonuclease Activity (Nicking):
The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
- Exonuclease Activity (Radioactivity Release):
The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
- Non-Specific DNase Activity (16 hour):
The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.
- Phosphatase Activity (PNPP):
The product is tested in a reaction containing a p-nitrophenyl phosphate (PNPP), a chromogenic substrate for most phosphatases. After incubation the percent degradation is determined by spectrophotometric analysis of released p-nitrophenol at 405 nm.
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Thermostable Inorganic Pyrophosphatase
- Heinonen, J.K. and Lahti, R.J. (1981). Analytical Biochemistry. 113, 313-317.
The supporting documents available for this product can be downloaded below.