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Blocking of targeted microRNAs from next-generation sequencing libraries.

ABSTRACT: 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.

SUBMITTER: Roberts BS

PROVIDER: S-EPMC4666382 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

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Blocking of targeted microRNAs from next-generation sequencing libraries.

Roberts Brian S BS Hardigan Andrew A AA Kirby Marie K MK Fitz-Gerald Meredith B MB Wilcox C Mel CM Kimberly Robert P RP Myers Richard M RM

Nucleic acids research 20150723 21

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 abun ...[more]

PMID: 26209131

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