Ribonuclease A is used to isolate DNA for RNA-free DNA. Ribonuclease A hydrolyzes RNA at C and U residues, cleaving between the 3’-Phosphate group of a pyrimidine nucleotide and the 5’-hydroxyl group of the adjacent nucleotide, i.e., those cleaving cyclic 2',3'-pyrimidine nucleotide residues either singly or at the termination of purine nucleotide chains (Volkin and Cohn 1953). The composition and structure of ribonuclease A has been extensively investigated. The tertiary structure was reported by Kartha, et al. (1967) and Lin, (1970). Wang, et al. (1975) indicate that flexibility in a three-dimensional structure is essential for optimal catalytic activity. Ribonuclease is inhibited by heavy metal ions and is competitively inhibited by DNA. The effect of denatured DNA is much greater than that of the native nucleic acid (Sekine et al. 1969).
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