Pyrophosphatase Inorganic(yeast) HYJ221
Inorganic pyrophosphatase (yeast) is derived from a recombinant Escherichia coli strain carrying the Saccharomyces cerevisiae inorganic pyrophosphatase gene. This enzyme catalyzes the hydrolysis of inorganic pyrophosphate to produce orthophosphate:
P207–4 + H20 → 2HP04–2
In reactions such as nucleic acid amplification and in vitro transcription, inorganic pyrophosphatase hydrolyzes the inorganic pyrophosphate generated during these reactions, thereby relieving the inhibitory effect of the generated inorganic pyrophosphate on the reaction system. The removal of pyrophosphate shifts the reaction equilibrium toward product formation, thereby increasing the yield of the synthesized product. For example, in RNA and DNA synthesis reactions, the action of inorganic pyrophosphatase shifts the reaction equilibrium toward product formation.
Components
|
Components |
10U |
50U |
100U |
1KU |
10KU |
|
Pyrophosphatase Inorganic(yeast) 0.1U/μL |
0.1 mL |
0.5 mL |
1 mL |
10 mL |
100 mL |
Storage
Store at -25 to -15°C. Valid for 2 years (avoid repeated freezing and thawing).
Storage buffer
20 mM Tris-HCl (pH 8.0), 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 50%glycerol.
Activity Definition
One activity unit (U) is defined as the amount of enzyme required to catalyze the hydrolysis of inorganic pyrophosphate to produce 1 μmol of phosphate per minute under standard reaction conditions.
Standard reaction conditions are: a 500 μL reaction system containing 100 mM Tris-HCl (pH 7.2, 25°C), 2 mM MgCl2, and 2 mM inorganic pyrophosphate, at 25°C for 10 min.
Quality Control
1.Exonuclease activity: When 0.1 U of this product was reacted with 1 μg of λ-Hind III digested DNA at 37°C for 16 hours, the electrophoretic band of the DNA remained unchanged.
2.Endonuclease activity: When 0.1 U of this product was reacted with 1 μg of λ DNA at 37°C for 16 hours, the electrophoretic band of the DNA remained unchanged.
3.Nickase activity: When 0.1 U of this product was reacted with 1 μg of pBR322 at 37°C for 16 hours, the electrophoretic band of the DNA remained unchanged.
4.RNase activity: When 0.1 U of this product was reacted with 1.6 μg of MS2 RNA at 37°C for 4 hours, the electrophoretic band of the RNA remained unchanged.
5.Residual E. coli DNA detection: Residual nucleic acid in 0.1 U of this product was detected by qPCR using an E. coli 16s rDNA-specific TaqMan PCR kit. Detection: E. coli genome residue ≤ 1 copy.
6.Endotoxin: LAL-Test, Chinese Pharmacopoeia 2020 Edition, Part IV, Gel Limit Test, General Rules(1143), bacterial endotoxin content ≤ 10 EU/mg.
Applications
Increase RNA yield in transcription reactions.