It cleaves the phosphodiester bond between the 5′-ribose of a nucleotide and the phosphate group attached to the 3′-ribose of an adjacent pyrimidine nucleotide. The resulting 2′, 3′-cyclic phosphate is hydrolyzed to the corresponding 3′-nucleoside phosphate.

Highlights

• RNase A is free of DNase activity. It is not necessary to heat it before use.

Applications

• Plasmid and genomic DNA preparation
• Removal of RNA from recombinant protein preparations
• Ribonuclease protection assays. Used in conjunction with RNase T1
• Mapping single-base mutations in DNA or RNA

Notes

Recommended concentration of RNase A is 1 to 100 µg/mL depending on the application. 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.