HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ANALYSIS OF LOW-MOLECULAR-MASS PRODUCTS SYNTHESIZED BY POLYNUCLEOTIDE PHOSPHORYLASE IN POLYMERIZATION REACTION

HPLC was used for the simultaneous analysis of low- and high-molecular -mass products of nucleotide polymerization. Oligonucleotides with cha in lengths of less than 5 monomer units were found in the reaction mix ture at the very beginning of the reaction. These oligomers were not t he products of non-specific polymer degradation. An increase in NaCl c oncentration from 0 to 1 IM resulted in the appearance of a short lag period and the oligonucleotide accumulation. A simultaneous increase i n Mg2+ concentration eliminated the lag. It is concluded that high ion ic strength affects both the formation of the metal-monomer-polymer co mplex and the affinity of the polynucleotide phosphorylase molecule fo r the reaction products.