Oligonucleotide Primers with G8AE-Clamp Modifications for RT-qPCR Detection of the Low-Copy dsRNA

Methods Mol Biol. 2019:1973:281-297. doi: 10.1007/978-1-4939-9216-4_18.

Abstract

We developed a new technique suitable for improved detection of low-copy dsRNA using modified oligonucleotides as primers in RT-qPCR. Insertion of G8AE-clamp residues into primers significantly improves thermal stability of duplexes with RNA without decrease of hybridization selectivity. The applicability of modified primers is demonstrated for detection of low-copy Kemerovo virus dsRNA.

Keywords: G-clamp; Modified primer; Oligonucleotide; RT-qPCR; dsRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Primers / chemistry*
  • Oligonucleotides / chemistry*
  • RNA, Double-Stranded / analysis*
  • RNA, Double-Stranded / chemistry
  • RNA, Double-Stranded / genetics
  • RNA, Viral / analysis*
  • RNA, Viral / chemistry
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods*

Substances

  • DNA Primers
  • Oligonucleotides
  • RNA, Double-Stranded
  • RNA, Viral