Description:
Inorganic pyrophosphatase (PPase) catalyzes the hydrolysis of inorganic pyrophosphate to form orthophosphate. It retains 100% activity after incubation at 100°C for 4 hours.
Application:
- Optimizes PCR through the elimination of pyrophosphate
- Catalyzes the conversion of inorganic pyrophosphate to orthophosphate
- Removes inhibiting amounts of pyrophosphates in the reaction
Source:
An E. coli strain carrying a plasmid encoding pyrophosphatase from the extreme thermophile Thermococcus litoralis.
Unit Definition:
One 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).
Buffer:
20 mM Tris-HCl
100 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
pH 8.0 @ 25°C
Recommended Storage Condition: -20°C
Reference:
Heinonen, J.K. and Lahti, R.J. (1981) Analytical Biochemistry, 113, 313-317.