Quantification of microRNAs by a simple and specific qPCR method

Susanna Cirera, Peter Kamp Busk

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14 Citationer (Scopus)

Abstrakt

MicroRNAs are powerful regulators of gene expression at post-transcriptional level and play important roles in many biological processes and in disease. The rapid pace of the emerging field of microRNAs has open new avenues for development of techniques to quantitatively determine microRNA expression levels in different systems.
In this chapter we describe a PCR method for quantification of microRNAs based on a single reverse transcription reaction for all microRNAs combined with real-time PCR with two microRNA-specific DNA primers. This method quantifies synthetic templates over eight orders of magnitude and successfully discriminate microRNAs that differ by one single nucleotide. Due to the usage of DNA primers this method allows higher amplification efficiencies than a similar method based on locked nucleic acids-spiked primers. The high efficiency translates into higher sensitivity and precision in microRNA quantification. Furthermore, the method is easy to perform with common laboratory reagents, which allows microRNA quantification at low cost.
OriginalsprogEngelsk
TitelRNA Mapping : Methods and Protocols
RedaktørerM. Lucrecia Alvarez, Mahtab Nourbakhsh
Antal sider9
Udgivelses stedNew York
ForlagSpringer Science+Business Media
Publikationsdato31 jul. 2014
Sider73-81
Kapitel7
ISBN (Trykt)978-1-4939-1061-8
ISBN (Elektronisk)978-1-4939-1062-5
DOI
StatusUdgivet - 31 jul. 2014
NavnMethods in Molecular Biology
Vol/bind1182
ISSN1064-3745

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Citationsformater

Cirera, S., & Busk, P. K. (2014). Quantification of microRNAs by a simple and specific qPCR method. I M. Lucrecia Alvarez, & M. Nourbakhsh (red.), RNA Mapping: Methods and Protocols (s. 73-81). Springer Science+Business Media. Methods in Molecular Biology, Bind. 1182 https://doi.org/10.1007/978-1-4939-1062-5_7