Tag/hybridization-based sensitive detection of polymerase chain reaction products

Kousuke Niwa, Akinobu Oribe, Hidemasa Okumura, Masahiro Shimono, Kenkichi Nagai, Toshikazu Hirota, Hiroshi Yasue, Mitsuo Kawase

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

The polymerase chain reaction (PCR) is an important technology to amplify a single copy or a few copies of DNA segment in genomic DNAs, visualizing the segment as DNA fragment. Thus, PCR is frequently used in various examinations such as detection of bacteria and fungi in the food industry. Here, we report a simple and sensitive method for detection of PCR products using single-strand tag sequence and hybridization of the tag sequence to the complementary tag sequence immobilized on solid material (STH). The detection sensitivity was found to be at least 50 times higher than electrophoresis/ethidium bromide (EtBr) visualization for approximately a 500-bp fragment and higher than the ordinary hybridization, that is, hybridization of denatured PCR product to probe sequence immobilized on solid material.

Original languageEnglish
Pages (from-to)12-16
Number of pages5
JournalAnalytical Biochemistry
Volume464
DOIs
Publication statusPublished - 2014 Nov 1

Keywords

  • DNA microarray
  • Fluorescence label
  • Hybridization
  • Non-nucleic acid spacer
  • Single-stranded DNA tag

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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