The sequential enzymatic determination of DNA and RNA

摘要: Abstract Enzymatically small quantities of native DNA and RNA in crude tissue homogenates can readily be specifically determined using ethidium bromide as a fluorescent indicator. The bromide-polynucleotide complexes serve effective substrates for deoxyribonuclease ribonuclease, respectively. Optimal divalent metal cation requirements were common reaction medium that is compatible both the DNase RNase reactions. To single mixture, contains biological sample, sequential addition produces specific rapid decrease fluorescence proportional to respective amounts present. Levels found six different tissues rat enzymatically by this method compared obtained alternate techniques. values highly reproducible correlate well with those more time-consuming, conventional methods. Most levels tissues, however, significantly greater than spectrophotometrically. Advantages enzymatic procedure analysis polynucleotide content are: (1) determination within sample aliquot allowing maximum use available sample; (2) extensive fractionation extraction are not required; (3) it especially useful when limited; (4) sensitivity 0.05 0.25 μg RNA, Reaction conditions developed assay also provide sensitive means continuously monitor hydrolysis; activity directly amount enzyme added.