RNase A, DNase and protease-free

Thermo Scientific RNase A, DNase and protease-free is an endoribonuclease that specifically degrades single-stranded RNA at C and U residues (see Figure 1 in Supporting Data).

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 (see References 1, 2).

Highlights

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

Applications

• Plasmid and genomic DNA preparation (see References 3, 4)
• Removal of RNA from recombinant protein preparations
• Ribonuclease protection assays. Used in conjunction with RNase T1 (see Reference 3)
• Mapping single-base mutations in DNA or RNA (see References 5, 6)

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 (see Reference 9).