RNase HII is a ribonuclease internal enzyme obtained from the Pyrococcus abyssi and recombinantly expressed in E. Coli. It recognizes the DNA-rN-DNA/DNA double strand, cutting at the site where ribonucleotides are incorporated into DNA. However, its activity on single-stranded RNA is very low, and it shows no cutting activity on dsDNA or ssDNA. RNase HII cuts at the site of a single ribonucleotide residue from the 5' end, producing a 5' phosphate group and a 3' hydroxyl end after cutting. This RNase HII enzyme has optimal activity at 70~75°C, is active between 50°C and 75°C, but has very low activity at room temperature. The product is highly thermostable, with almost no loss of activity after incubation at 95°C for 45 minutes, and is compatible with various PCR reaction systems. Specifications Cat.No. 14539ES80/14539ES92 Size 50 U / 250 U Species Pyrococcus abyssi, P.a. Activity One unit is defined as the amount of enzyme necessary to nick 1 nmol of synthetic, single-rC-containing duplex substrate per minute at 70°C in 10 mM Tris, 50 mM NaCl, 0.01% Triton X-100, 10 µg/ml BSA, 4 mM MgCl2. Depending on the application, enzyme usage range is 2−200 mU per reaction. Components Name 14539ES50 (50U) 14539ES72 (250U) RNase HII（2 U/μL） 25 μL 125 μL Storage This product should be stored at-25~-15℃ for 2 years. Product Application Detection by LAMP with high-sensitivity probes. RNase HII-dependent PCR (rhPCR). Removal of mismatched ribonucleotides formed during polymerase chain reaction. Degradation of the RNA portion of Okazaki fragments. Notes This product is for research use only. Please operate with lab coats and disposable gloves, for your safety. Figures 1. E. coli genomic DNA residue < 0.5 copies/100 U Detection of E. coli genomic DNA residue in different batches of RNase HII showed that the host genomic DNA residue of YEASEN's RNase HII is far below 0.5 copies/100 U. Fig 4. Detection of E. coli genomic DNA residue 2. 95°C Heat Resistance Test After heating RNase HII at 95°C for 0 to 45 minutes, the enzyme activity of RNase HII was tested. The results indicated that there was almost no loss of enzyme activity in YEASEN's RNase HII even after 45 minutes of heating. Fig 5. 95°C heat resistance test 3. No Exonuclease., Nicking Enzyme, or RNase Residue (1 U Dose) Incubating 1 U of different batches of RNase HII with their respective substrates DNA/RNA at 37°C for 4 hours, the results of agarose gel electrophoresis indicated that there was no exonuclease, nicking enzyme, or RNase residue in RNase HII. Fig 6. Detection of exonuclease, nicking enzyme, and RNase residue Documents: Manuals
