Rapid DNA chemical ligation for amplification of RNA and DNA signal

Hiroshi Abe, Yuko Kondo, Hiroshi Jinmei, Naoko Abe, Kazuhiro Furukawa, Atsushi Uchiyama, Satoshi Tsuneda, Kyoko Aikawa, Isamu Matsumoto, Yoshihiro Ito

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Enzymatic ligation methods are useful in the diagnostic detection of DNA sequences. Here, we describe the investigation of nonenzymatic phosphorothioate-iodoacetyl DNA chemical ligation as a method for the detection and identification of RNA and DNA. The specificity of ligation on the DNA target is shown to allow the discrimination of a single point mutation with a drop in the ligation yield of up to 16.1-fold. Although enzymatic ligation has very low activity for RNA targets, this reaction is very efficient for RNA targets. The speed of the chemical ligation with an RNA target achieves a 70% yield in 5 s, which is equal to or better than that of ligase-enzyme-mediated ligation with a DNA target. The reaction also exhibits a significant level of signal amplification under thermal cycling in periods as short as 100-120 min, with the RNA or DNA target acting in a catalytic way to ligate multiple pairs of probes.

Original languageEnglish
Pages (from-to)327-333
Number of pages7
JournalBioconjugate Chemistry
Volume19
Issue number1
DOIs
Publication statusPublished - 2008 Jan 1

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

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  • Cite this

    Abe, H., Kondo, Y., Jinmei, H., Abe, N., Furukawa, K., Uchiyama, A., Tsuneda, S., Aikawa, K., Matsumoto, I., & Ito, Y. (2008). Rapid DNA chemical ligation for amplification of RNA and DNA signal. Bioconjugate Chemistry, 19(1), 327-333. https://doi.org/10.1021/bc700244s