RNase A MBG

• Possesses enzyme activity of >80 units/mg protein
• Degrades RNA to cyclic nucleotide monophosphates to 5’-OH and 2’-, 3’-cyclic monophosphate
• No endonuclease or exonuclease activity towards DNA substrates
• Selectively cleaves single-stranded RNA 3’ next to pyrimidine residues

The Ribonuclease A (RNase A) is a 13.7 kDa (monomer) endoribonuclease isolated from bovine pancreas, which selectively cleaves single-stranded RNA 3’ next to pyrimidine residues (cytosine, uracil). It degrades RNA to cyclic nucleotide monophosphates to 5’- OH and 2’-, 3’-cyclic monophosphate. The enzyme exhibits no endonuclease or exonuclease activity toward DNA substrates. RNase A removes RNA during the isolation procedures of plasmid and genomic DNA.

It is supplied with 1–10 mg/mL by resuspending in 10 mM Tris-HCl (pH 7.5), 15 mM NaCl, 50% (v/v) glycerol or in TE buffer.

One unit of activity is defined as the amount of enzyme which causes the hydrolysis of RNA to yield a velocity constant, k=1, at 25°C and pH 5.0.

RNase A is very active under a wide range of reaction conditions and is difficult to inactivate. At low salt concentrations (up to 100 mM NaCl), the RNase A cleaves single- and double-stranded RNA as well as an RNA strand in RNA-DNA hybrids. However, under high salt concentrations (>300 mM NaCl), RNase A specifically cleaves single-stranded RNA.

RNase A cleaves specifically at the 3′-side of pyrimidine (uracil or cytosine) phosphate bonds. Ribonucleases do not hydrolyze DNA because the DNA lacks 2′-OH groups essential for forming cyclic intermediates. The enzyme is active under a wide range of reaction conditions. At low salt concentrations (0 to 100 mM NaCl), RNase A cleaves single-stranded and double-stranded RNA as well the RNA strand in RNA-DNA hybrids. However, at NaCl concentrations of 0.3 M or higher, RNase A specifically cleaves single-stranded RNA. RNase is supplied as a lyophilized powder.

Usage

Stock solutions should be prepared to a final concentration of 1–10 mg/mL by resuspending in 10 mM Tris-HCl (pH 7.5), 15 mM NaCl, 50% (v/v) glycerol or in TE buffer. 

The recommended working solution concentration depends on the application.

• For removal of RNA from plasmid preparations, use 10 μg/mL working solution and incubate the sample for 1 hour at room temperature.
• For the preparation of „blunt ends” of double-stranded cDNA, use a 100 ng/mL working solution.

This is used for applications such as:

• Purification of RNA-free DNA
• Isolation of plasmid and genomic DNA
• Removal of RNA during recombinant protein preparations
• Protection of RNA during assays
• Mapping of single-based mutations in DNA or RNA

The acquisition of BLIRT by QIAGEN in the second quarter of 2022 paved the way for BLIRT’s expansion of R&D and manufacturing capabilities for customized and standardized recombinant enzymes and molecular biology reagents. Due to the transition, the rebranding process from BLIRT to QIAGEN is ongoing and the following resources still show BLIRT branding.
 
We assure you that the integration does not affect the quality, fit, form and function of the products.