Open AccessA simple method for the preparation of (β-32P)purine nucleoside triphosphates
Author(s) -
Yasuhiro Furuichi,
Aaron J. Shatkin
Publication year - 1977
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/4.10.3341
Subject(s) - deoxyribonucleoside , guanosine , adenylate kinase , nucleoside , biochemistry , biology , ribonucleoside , nucleoside diphosphate kinase , purine , nucleotide , pyruvate kinase , enzyme , purine nucleoside phosphorylase , guanine , guanosine triphosphate , gtp' , rna , glycolysis , gene
A rapid, simple and inexpensive procedure is described for the preparation of purine ribo-and deoxyribonucleoside triphosphates specifically and highly radiolabeled with [32P]phosphate in the beta position. The method involves two successive enzymatic reactions: conversion of donor [gamma-32P]ATP in the presence of an excess of acceptor 5'-mononucleotide to the 5'-diphosphates by myokinase or guanosine 5'-monophosphate kinase followed by phosphorylation with pyruvate kinase to yield 5'-triphosphates.