Blocking of targeted microRNAs from next-generation sequencing libraries

Academic Article

Abstract

  • © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. Highly abundant microRNAs (miRNAs) in small RNA sequencing libraries make it difficult to obtain efficient measurements of more lowly expressed species. We present a new method that allows for the selective blocking of specific, abundant miRNAs during preparation of sequencing libraries. This technique is specific with little off-target effects and has no impact on the reproducibility of the measurement of non-targeted species. In human plasma samples, we demonstrate that blocking of highly abundant hsa-miR-16-5p leads to improved detection of lowly expressed miRNAs and more precise measurement of differential expression overall. Furthermore, we establish the ability to target a second abundant miRNA and to multiplex the blocking of two miRNAs simultaneously. For small RNA sequencing, this technique could fill a similar role as do ribosomal or globin removal technologies in messenger RNA sequencing.
  • Published In

    Digital Object Identifier (doi)

    Author List

  • Roberts BS; Hardigan AA; Kirby MK; Fitz-Gerald MB; Wilcox CM; Kimberly RP; Myers RM
  • Volume

  • 43
  • Issue

  • 21