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Comparison of methods for miRNA isolation and quantification from ovine plasma.

ABSTRACT: microRNA (miRNA) are promising candidates for disease biomarkers as they are abundant in circulation, highly stable in biological fluids and may yield diagnostic biomarker signatures. The reported issues with miRNA isolation using traditional RNA reagents necessitates the optimisation of miRNA isolation from challenging samples. In this study we compared six commercial RNA extraction kits to evaluate their ability to isolate miRNA from ovine plasma. We also compared three methods for quantification of small RNA extracted from plasma to determine the most reliable. Using minimal sample inputs of fresh and frozen plasma from five sheep, we compared the six kits (Kit A-F) using quantitative PCR. Operational factors were also assessed for each kit. Kits A and B provided the best detection of the miRNA qPCR reference genes across fresh and frozen samples (p?

SUBMITTER: Wright K

PROVIDER: S-EPMC6972740 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Comparison of methods for miRNA isolation and quantification from ovine plasma.

Wright Kathryn K de Silva Kumudika K Purdie Auriol C AC Plain Karren M KM

Scientific reports 20200121 1

microRNA (miRNA) are promising candidates for disease biomarkers as they are abundant in circulation, highly stable in biological fluids and may yield diagnostic biomarker signatures. The reported issues with miRNA isolation using traditional RNA reagents necessitates the optimisation of miRNA isolation from challenging samples. In this study we compared six commercial RNA extraction kits to evaluate their ability to isolate miRNA from ovine plasma. We also compared three methods for quantificat ...[more]

PMID: 31964966

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