Self-reporting hybridisation assay for miRNA analysis

Jo-Anne Riley, Tom Brown, Nittaya Gale, Julie Herniman, G. John Langley*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)
4 Downloads (Pure)

Abstract

Hybridisation assays, which are commonly used to analyse oligonucleotides such as siRNAs and miRNAs, often employ detection probes with fluorescent tags. The signal emitted by a fluorescent tag covers a broad range of wavelengths and this limits the multiplexing potential due to overlapping signals. A novel method of indirect oligonucleotide analysis has been developed which combines a hybridisation assay with cleavable small molecule mass tags using HPLC-ESI MS detection. A self-reporting detection probe has been designed which incorporates a DNA/RNA chimeric oligonucleotide sequence in the reporter region, which generates small nucleotide products upon RNase cleavage of the ribose-phosphate backbone. These small nucleotides can then serve as mass tags for the indirect detection of oligonucleotide analytes. The narrow mass range covered by a small molecule mass tag combined with the wide range of possible mass tags provides a high degree of multiplexing potential. This approach has been demonstrated for the analysis of a synthetic miRNA.

Original languageEnglish
Pages (from-to)1088-1092
Number of pages5
JournalAnalyst
Volume139
Issue number5
DOIs
Publication statusPublished - 7 Mar 2014
Externally publishedYes

Bibliographical note

Copyright the Publisher 2014. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • NUCLEIC-ACIDS
  • MICRORNA
  • PROBES
  • RNA
  • ELEGANS

Cite this