Activation of DNA primer for DNA polymerase as an ultrasensitive assay for a subgroup of enzymes with deoxyribonuclease activity

George P. Studzinski, Gary J. Fischman

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Pretreatment of native DNA with nucleases which hydrolyze phosphodiester bonds with liberation of 3′-hydroxyl terminal groups is known to increase the rate of incorporation of nucleotides into DNA by E. coli DNA polymerase I and similar enzymes. Concentration ranges and conditions for this reaction have been established which allow specific detection and quantitative assay of such nucleases. The method permits detection of as little as 0.1 pg of pancreatic deoxyribonuclease, is simple, and requires only commercially available components. It has been utilized to monitor purification of a nuclease from HeLa cell nuclei at concentrations which could not be detected by other methods.

Original languageEnglish (US)
Pages (from-to)449-458
Number of pages10
JournalAnalytical Biochemistry
Volume58
Issue number2
DOIs
StatePublished - Apr 1974
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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