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Cat. No.: ETRHII-200 (for 200U)

Cat. No.: ETRHII-1k (for 1KU)

Description

Extreme Thermostable RNase HII is an endogenous endonuclease derived from extremely thermostable strains. It recognizes RNA/DNA hybrid strands and cleaves RNA strands. It has very low cleavage activity to single-stranded RNA and has no cleavage activity to dsDNA and ssDNA. The RNase HII enzyme cleaves from the 5' end of the single ribonucleotide residue of DNA/RNA duplex to produce a 5' - terminal phosphate group and a 3 '- terminal hydroxyl group. The RNase HII enzyme has the best activity at 70-75°C and is active at 50°C to 75°C, but its activity is very low at room temperature (activity is reduced by~1000 times). The enzyme is extremely heat-resistant and retains all its activity after being heated at 95°C for 15 minutes. Therefore, it is compatible with most PCR buffer systems and can be used in RNase HII dependent PCR (rhPCR) and other experiments. Compared with Rnase HI, Rnase HII can recognize and cut single ribonucleotides, while RNase HI recognizes 2 '- OH of four consecutive ribonucleotides; In addition, Rnase HII can degrade Okazaki fragments, while RNase HI has no such activity.

Unit Definition

One unit is defined as the amount of enzyme required to cut 1 nmol of synthetic DNA/RNA hybrid double-stranded substrate containing single rC per minute in 10 mM Tris, 50 mM NaCl, 0.01% Triton X-100, 10 µg/ml BSA, and 4 mM MgCl₂ at 70°C.

Components

200U

ET RNase HII (2U/μl): 100 μl

Applications

• Reduce or remove primer dimer in PCR reaction

• SNP detection and rare allele detection

• Differentiate heterologous genes

• Improve the accuracy of multiplex PCR products

Storage

The minimum shelf life is 3 years at -20°C.

Note

• When this enzyme is used in rhPCR (RHase HII dependent PCR), it needs to use a DNA primer with a 3' end closure group.
• The enzyme is active at 50-75°C, and its dosage can be adjusted according to different applications.
• The enzyme is compatible with most PCR buffer systems and has broad requirements for Mg²⁺concentration. It can play an active role between 1~6 mM.
• The enzyme is extremely heat-resistant, and its activity does not decrease significantly at 95°C for 15 minutes, so it cannot be inactivated by heat, but can be inactivated by 0.1% SDS.