Substantial background reduction in ligase-based apoptosis detection using newly designed hairpin oligonucleotide probes

Biotechniques. 1999 Dec;27(6):1130-2. doi: 10.2144/99276bm09.

Abstract

A method for selective detection of apoptotic cells by ligation of DNA probes to the ends of specific double-stranded DNA breaks in tissue sections was recently reported (1). Unfortunately, this method produces high levels of background staining, due to nonspecific attachment of the probes to the tissue sections, and thereby requires extensive and repeated washing. In the current study we redesigned the oligoprobes in a manner that virtually eliminated background staining. This new design substantially reduces the cost of the probe preparation, making it a convenient and robust methodology to detect apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Apoptosis / genetics*
  • Base Sequence
  • Biotinylation
  • DNA / analysis
  • DNA / genetics
  • DNA Fragmentation / genetics
  • Dexamethasone / pharmacology
  • Ligases / metabolism*
  • Nucleic Acid Conformation
  • Oligonucleotide Probes / chemistry
  • Oligonucleotide Probes / genetics*
  • Rats
  • Sensitivity and Specificity
  • Thymus Gland / cytology
  • Thymus Gland / drug effects
  • Thymus Gland / metabolism

Substances

  • Oligonucleotide Probes
  • Dexamethasone
  • DNA
  • Ligases