Pancreatic RNase A specifically cleaves at the 3′-side of pyrimidine (uracil or cytosine) phosphate bonds. The enzyme does not hydrolyze DNA, because DNA lacks 2′-OH groups essential for the formation of cyclic intermediates. The enzyme can also be used to hydrolyze RNA from protein samples. It is compatible for use in RNase protection assays, to remove unspecifically bound RNA, in the analysis of RNA sequences, to hydrolyze RNA contained in protein samples, and in the purification of DNA.