Abstract
Reverse-transcriptase quantitative PCR (RT-qPCR) is a widely used method for quantifying microRNAs (miRNAs) in cells and tissues. However, the quantification of miRNAs in the circulation presents specific challenges. Here, we describe an optimized protocol using a small amount of input material to assess serum sample quality and quantify levels of a panel of up to 20 miRNAs. This is achieved by multiplexing Taqman miRNA stem-loop primers in the reverse transcription step. An additional multiplexed pre-amplification step is used to increase the sensitivity of the final quantification step, which is carried out using standard Taqman qPCR methodology.
*Joint first authors.
† Joint senior authors.
†Joint senior authors.
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Bell, E., Watson, H.L., Bailey, S., Murray, M.J., Coleman, N. (2017). A Robust Protocol to Quantify Circulating Cancer Biomarker MicroRNAs. In: Dalmay, T. (eds) MicroRNA Detection and Target Identification. Methods in Molecular Biology, vol 1580. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6866-4_18
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DOI: https://doi.org/10.1007/978-1-4939-6866-4_18
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-6866-4
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