A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity

Research output: Contribution to journalArticlepeer-review

Abstract

A technique for conveniently radiolabeling DNA restriction endonuclease fragments to high specific activity is described. DNA fragments are purified from agarose gels directly by ethanol precipitation and are then denatured and labeled with the large fragment of DNA polymerase I, using random oligonucleotides as primers. Over 70% of the precursor triphosphate is routinely incorporated into complementary DNA, and specific activities of over 109 dpm/μg of DNA can be obtained using relatively small amounts of precursor. These "oligolabeled" DNA fragments serve as efficient probes in filter hybridization experiments.

Original languageEnglish (US)
Pages (from-to)6-13
Number of pages8
JournalAnalytical biochemistry
Volume132
Issue number1
DOIs
StatePublished - Jul 1 1983

Keywords

  • DNA fragments
  • Southern transfers
  • agarose gels
  • cloned DNA
  • hybridization
  • radiolabeling

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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