Stem-Loop qRT-PCR-Based Quantification of miRNAs

Yoann Abel; Mathieu Rederstorff

doi:10.1007/978-1-0716-1386-3_6

Stem-Loop qRT-PCR-Based Quantification of miRNAs

Methods Mol Biol. 2021:2300:59-64. doi: 10.1007/978-1-0716-1386-3_6.

Authors

Yoann Abel¹, Mathieu Rederstorff²

Affiliations

¹ IGMM, CNRS, Université de Montpellier, Montpellier, France. yoann.abel@igmm.cnrs.fr.
² Université de Lorraine, CNRS, IMoPA, Nancy, France.

PMID: 33792871
DOI: 10.1007/978-1-0716-1386-3_6

Abstract

Stem-loop qRT-PCR is one of the most commonly used real-time PCR approach to quantify small non-coding RNAs such as microRNAs. The quantification method is divided in two steps. First, RNA is reverse transcribed using a specific stem-loop primer, and the resulting RT product is subsequently used as a template for quantitative real-time PCR. This fast and simple method provides quantitative data with high sensitivity and specificity to study miRNAs and their functions.

Keywords: MicroRNA; Real-time PCR quantification; Small non-coding RNA; Stem-loop primers.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Inverted Repeat Sequences
MicroRNAs / analysis*
MicroRNAs / chemistry*
Real-Time Polymerase Chain Reaction
Reverse Transcription

Substances

MicroRNAs