Quantification of microRNAs by a simple and specific qPCR method

Susanna Cirera, Peter Kamp Busk

Research output: Contribution to book/anthology/report/conference proceedingBook chapterResearchpeer-review

27 Citations (Scopus)

Abstract

MicroRNAs (miRNAs) are powerful regulators of gene expression at posttranscriptional level and play important roles in many biological processes and in disease. The rapid pace of the emerging field of miRNAs has opened new avenues for development of techniques to quantitatively determine miRNA expression levels in different systems. In this chapter we describe a PCR method for quantification of miRNAs based on a single reverse transcription reaction for all miRNAs combined with real-time PCR with two miRNA-specific DNA primers. This method quantifies synthetic templates over eight orders of magnitude and successfully discriminates miRNAs 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 acid-spiked primers. The high efficiency translates into higher sensitivity and precision in miRNA quantification. Furthermore, the method is easy to perform with common laboratory reagents, which allows miRNA quantification at low cost.
Original languageEnglish
Title of host publicationRNA Mapping : Methods and Protocols
EditorsM. Lucrecia Alvarez, Mahtab Nourbakhsh
Number of pages9
Place of PublicationNew York
PublisherSpringer Science+Business Media
Publication date31 Jul 2014
Pages73-81
Chapter7
ISBN (Print)978-1-4939-1061-8
ISBN (Electronic)978-1-4939-1062-5
DOIs
Publication statusPublished - 31 Jul 2014
SeriesMethods in Molecular Biology
Volume1182
ISSN1064-3745

Fingerprint

Dive into the research topics of 'Quantification of microRNAs by a simple and specific qPCR method'. Together they form a unique fingerprint.

Cite this