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Anal Chem. 2015 Jan 20;87(2):1404-10. doi: 10.1021/ac504406s. Epub 2014 Dec 26.

Highly-sensitive amplification-free analysis of multiple miRNAs by capillary electrophoresis.

Author information

1
Department of Chemistry and Centre for Research on Biomolecular Interactions, York University , 4700 Keele Street, Toronto, Ontario M3J 1P3, Canada.

Abstract

Sets of deregulated microRNAs (miRNAs), termed miRNA signatures, are promising biomarkers for cancer. Validation of miRNA signatures requires a technique that is accurate, sensitive, capable of detecting multiple miRNAs, fast, robust, and not cost-prohibitive. Direct quantitative analysis of multiple miRNAs (DQAMmiR) is a capillary electrophoresis (CE)-based hybridization assay that was suggested as a methodological platform for validation and clinical use of miRNA signatures. While satisfying the other requirements, DQAMmiR is not sufficiently sensitive to detect low-abundance miRNAs. Here, we solve this problem by combining DQAMmiR with the preconcentration technique, isotachophoresis (ITP). The sensitivity improved 100 times (to 1 pM) allowing us to detect low-abundance miRNAs in an RNA extract. Importantly, ITP-DQAMmiR can be performed in a fully automated mode using a commercial CE instrument making it suitable for practical applications.

PMID:
25495883
DOI:
10.1021/ac504406s
[Indexed for MEDLINE]

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