MicroRNA Analysis Using the Quantitative Real-Time PCR Reaction

Methods Mol Biol. 2018:1823:69-85. doi: 10.1007/978-1-4939-8624-8_7.

Abstract

The analysis of microRNA expression patterns provides new insights into numerous cellular processes and their aberrances in diseases. Despite its potential pitfalls, the quantitative real-time polymerase chain reaction (qPCR) is the most commonly used tool for microRNA profiling. The method requires extraction and quality analysis of RNA, which is further reverse transcribed using specific primers and used as a template in a qPCR reaction. All these elements have been addressed in this chapter.

Keywords: FFPE samples; RNA extraction; Serum samples; Stem-loop primer; TaqMan probe; microRNA identification; microRNA quantification; qPCR.

MeSH terms

  • Gene Expression Profiling / methods*
  • MicroRNAs* / biosynthesis
  • MicroRNAs* / genetics
  • Real-Time Polymerase Chain Reaction / methods*

Substances

  • MicroRNAs