M.B. Enzymes GmbH

Tth inorganic pyrophosphatase.

Description

Tth inorganic pyrophosphatase (PPase, EC 3.6.1.1) catalyses the hydrolysis of inorganic pyrophosphate (which inhibit amplification reactions) to form orthophosphate: P₂O₇^-4+H₂O-> 2HPO₄^-2. The enzyme is extremely thermostable, retaining >90% activity after incubation at 100°C for 1 h.

Unit definition

1 unit will generate 40 nmol of phosphate per min from pyrophosphate under standard reaction conditions (50 mM Tricine pH 8.5, 1 mM MgCl₂, 0,32 mM PPi at 75°C for 10 min)

Reaction conditions

The enzyme works well in all known polymerase buffers. Titrating the amount of the enzyme used in standard amplification reaction can result in significant improvements in DNA amplification. Recommended range of enzyme concentration is 1-100 mU per reaction.

Storage

Store at -20°C.

Applications

maintains forward direction of reactions generating pyrophosphate (e.g., high temperature sequencing, long-template PCR); oligo-labeling; removing CAP structure on mRNA, viral RNA and low molecular weight RNA, by cleaving the connecting 5´-5´ triphosphate link; 5´- and 3´-end mapping of mRNA.

Quality control

activity, absence of ATPase and phosphatase activities.

References

1. Hohne, W.E. et al. (1988) Biomed Biochim Acta. 47, 941-947. 2. Satoh, T. et al. (1998) J. Biochem. (Tokyo) 124, 79-88.

Catalog #

T11.1

T11.2

Pack size

100 U

250 U

Tth inorganic pyrophosphatase.
Description	Tth inorganic pyrophosphatase (PPase, EC 3.6.1.1) catalyses the hydrolysis of inorganic pyrophosphate (which inhibit amplification reactions) to form orthophosphate: P₂O₇^-4+H₂O-> 2HPO₄^-2. The enzyme is extremely thermostable, retaining >90% activity after incubation at 100°C for 1 h.
Unit definition	1 unit will generate 40 nmol of phosphate per min from pyrophosphate under standard reaction conditions (50 mM Tricine pH 8.5, 1 mM MgCl₂, 0,32 mM PPi at 75°C for 10 min)
Reaction conditions	The enzyme works well in all known polymerase buffers. Titrating the amount of the enzyme used in standard amplification reaction can result in significant improvements in DNA amplification. Recommended range of enzyme concentration is 1-100 mU per reaction.
Storage	Store at -20°C.
Applications	maintains forward direction of reactions generating pyrophosphate (e.g., high temperature sequencing, long-template PCR); oligo-labeling; removing CAP structure on mRNA, viral RNA and low molecular weight RNA, by cleaving the connecting 5´-5´ triphosphate link; 5´- and 3´-end mapping of mRNA.
Quality control	activity, absence of ATPase and phosphatase activities.
References	1. Hohne, W.E. et al. (1988) Biomed Biochim Acta. 47, 941-947. 2. Satoh, T. et al. (1998) J. Biochem. (Tokyo) 124, 79-88.
Catalog #	T11.1	T11.2
Pack size	100 U	250 U