Triazole-linked DNA as a primer surrogate in the synthesis of first-strand cDNA

Tomoko Fujino, Ken Ichi Yasumoto, Naomi Yamazaki, Ai Hasome, Kazuhiro Sogawa, Hiroyuki Isobe

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

A phosphate-eliminated nonnatural oligonucleotide serves as a primer surrogate in reverse transcription reaction of mRNA. Despite of the nonnatural triazole linkages in the surrogate, the reverse transcriptase effectively elongated cDNA sequences on the 3'-downstream of the primer by transcription of the complementary sequence of mRNA. A structure-activity comparison with the reference natural oligonucleotides shows the superior priming activity of the surrogate containing triazole-linkages. The nonnatural linkages also protect the transcribed cDNA from digestion reactions with 5'-exonuclease and enable us to remove noise transcripts of unknown origins. A phosphate-eliminated surrogate of oligonucleotide is a primer substrate for reverse transcription of mRNA. The nonnatural linkages protect the transcribed cDNA from digestion reactions with 5'-exonuclease and enable us to remove noise transcripts of unknown origins.

Original languageEnglish
Pages (from-to)2956-2960
Number of pages5
JournalChemistry - An Asian Journal
Volume6
Issue number11
DOIs
Publication statusPublished - 2011 Nov 4

Keywords

  • DNA
  • enzymes
  • oligonucleotides
  • primers
  • reverse transcription

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry

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