MicroRNA Analysis Using the Quantitative Real-Time PCR Reaction
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.
KeywordsmicroRNA identification microRNA quantification TaqMan probe qPCR Stem-loop primer RNA extraction FFPE samples Serum samples
- 7.Wojcicka A, Swierniak M, Kornasiewicz O, Gierlikowski W, Maciag M, Kolanowska M et al (2014) Next generation sequencing reveals microRNA isoforms in liver cirrhosis and hepatocellular carcinoma. Int J Biochem Cell Biol 53:208–217. https://doi.org/10.1016/j.biocel.2014.05.020 CrossRefPubMedGoogle Scholar
- 8.Jazdzewski K, Liyanarachchi S, Swierniak M, Pachucki J, Ringel MD, Jarzab B et al (2009) Polymorphic mature microRNAs from passenger strand of pre-miR-146a contribute to thyroid cancer. Proc Natl Acad Sci U S A 106(5):1502–1505. https://doi.org/10.1073/pnas.0812591106 CrossRefPubMedPubMedCentralGoogle Scholar