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MicroRNA profiling in plasma samples using qPCR arrays: Recommendations for correct analysis and interpretation

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  • Andreas B Gevaert
  • Isabel Witvrouwen
  • Christiaan J Vrints
  • Hein Heidbuchel
  • Emeline M Van Craenenbroeck
  • Steven J Van Laere
  • Amaryllis H Van Craenenbroeck

Abstract

MicroRNA (miRNA) regulate gene expression through posttranscriptional mRNA degradation or suppression of translation. Many (pre)analytical issues remain to be resolved for miRNA screening with TaqMan Low Density Arrays (TLDA) in plasma samples, such as optimal RNA isolation, preamplification and data normalization. We optimized the TLDA protocol using three RNA isolation protocols and preamplification dilutions. By using 100μL elution volume during RNA isolation and adding a preamplification step without dilution, 49% of wells were amplified. Informative target miRNA were defined as having quantification cycle values ≤35 in at least 20% of samples and low technical variability (CV across 2 duplicates of 1 sample

Suggested Citation

  • Andreas B Gevaert & Isabel Witvrouwen & Christiaan J Vrints & Hein Heidbuchel & Emeline M Van Craenenbroeck & Steven J Van Laere & Amaryllis H Van Craenenbroeck, 2018. "MicroRNA profiling in plasma samples using qPCR arrays: Recommendations for correct analysis and interpretation," PLOS ONE, Public Library of Science, vol. 13(2), pages 1-13, February.
    • Handle: RePEc:plo:pone00:0193173
    DOI: 10.1371/journal.pone.0193173
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    References listed on IDEAS

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    1. Eric M. Small & Eric N. Olson, 2011. "Pervasive roles of microRNAs in cardiovascular biology," Nature, Nature, vol. 469(7330), pages 336-342, January.
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