[Preparation of fluorescent-labeled DNA and its use as a probe in molecular hybridization]

Bioorg Khim. 1986 Nov;12(11):1508-13.
[Article in Russian]

Abstract

A method of the fluorescent-labeled DNA preparation for visualization of the complementary nucleotide sequences has been developed. Polynucleotide probes were alkylated randomly by 4-(N-methylamino-N-2-chloroethyl)-benzylamine followed by modification with such fluorochromes as dansyl chloride or fluorescein isothiocyanate (FITC). It was found that the FITC but not dansyl-labeled polynucleotides could serve as efficient probes when about 4% of nitrogen bases were modified. The conditions minimizing the loss of the alkylated bases from DNA were determined. The procedure for hybridization with FITC-labeled DNA as a probe is described, concentration of DNA probe being about 4 ng/mm2 of the nitrocellulose filter.

Publication types

  • English Abstract

MeSH terms

  • Alkylation
  • DNA*
  • Fluorescent Dyes*
  • Nucleic Acid Hybridization*
  • Spectrometry, Fluorescence

Substances

  • Fluorescent Dyes
  • DNA