Reverse Transcription-Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) for Gene Expression Analyses

Albert Caballero-Solares; Jennifer R Hall; Xi Xue; Matthew L Rise

doi:10.1007/978-1-0716-2376-3_21

Reverse Transcription-Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) for Gene Expression Analyses

Methods Mol Biol. 2022:2508:319-340. doi: 10.1007/978-1-0716-2376-3_21.

Authors

Albert Caballero-Solares¹, Jennifer R Hall², Xi Xue³, Matthew L Rise³

Affiliations

¹ Department of Ocean Sciences, Memorial University of Newfoundland, St. John's, NL, Canada. acaballeroso@mun.ca.
² Aquatic Research Cluster, CREAIT Network, Ocean Sciences Centre, Memorial University of Newfoundland, St. John's, NL, Canada.
³ Department of Ocean Sciences, Memorial University of Newfoundland, St. John's, NL, Canada.

PMID: 35737248
DOI: 10.1007/978-1-0716-2376-3_21

Abstract

The reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) is considered to be the gold standard for gene expression research. However, for this claim to be valid, RT-qPCR studies must test and optimize the quality of its RNA templates and assays. This chapter describes the experimental procedures required to generate reliable and reproducible gene expression results using RT-qPCR.

Keywords: Gene of interest; MIQE guidelines; Normalizer gene selection; Polymerase chain reaction; Primer pairs; Quality testing; RNA; Reverse transcription; cDNA.

MeSH terms

RNA* / genetics
Real-Time Polymerase Chain Reaction / methods
Reverse Transcriptase Polymerase Chain Reaction
Reverse Transcription*

Substances

RNA