3'-End labeling of nucleic acids by a polymerase ribozyme

Nucleic Acids Res. 2018 Sep 28;46(17):e103. doi: 10.1093/nar/gky513.

Abstract

A polymerase ribozyme can be used to label the 3' end of RNA or DNA molecules by incorporating a variety of functionalized nucleotide analogs. Guided by a complementary template, the ribozyme adds a single nucleotide that may contain a fluorophore, biotin, azide or alkyne moiety, thus enabling the detection and/or capture of selectively labeled materials. Employing a variety of commercially available nucleotide analogs, efficient labeling was demonstrated for model RNAs and DNAs, human microRNAs and natural tRNA.

Publication types

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

MeSH terms

  • 3' Flanking Region*
  • Biotin / chemistry
  • Biotin / metabolism
  • DNA / chemistry
  • DNA / metabolism
  • DNA-Directed RNA Polymerases / metabolism*
  • Fluorescein / chemistry
  • Fluorescein / metabolism
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • MicroRNAs / chemistry
  • MicroRNAs / metabolism
  • Nucleic Acid Conformation
  • Nucleic Acids / chemistry
  • Nucleic Acids / metabolism*
  • Nucleotidyltransferases / metabolism
  • RNA / chemistry
  • RNA / metabolism
  • RNA, Catalytic / metabolism*
  • RNA, Transfer / chemistry
  • RNA, Transfer / metabolism
  • Rhodamines / chemistry
  • Rhodamines / metabolism
  • Staining and Labeling / methods*

Substances

  • Fluorescent Dyes
  • MicroRNAs
  • Nucleic Acids
  • RNA, Catalytic
  • Rhodamines
  • RNA
  • Biotin
  • DNA
  • RNA, Transfer
  • Nucleotidyltransferases
  • DNA-Directed RNA Polymerases
  • Fluorescein