Quantitative (Real-Time) RT-PCR in cardiovascular research

Kevin John Ashton, John Patrick Headrick

Research output: Chapter in Book/Report/Conference proceedingChapterResearchpeer-review

9 Citations (Scopus)

Abstract

Quantitative (real-time) PCR (qPCR) represents a highly sensitive, sequence-specific, and reproducible technique for the gel-free detection and quantitation of nucleic acids. Owing to its large dynamic range and throughput, this approach has become the chosen method for rapid quantification of mRNA levels in biological samples. The sensitivity of this method permits the reliable detection of low concentrations of initial template, while delivering a linear range of up to 10 orders of magnitude in copy number. This chapter details the basic methodology behind key components of a qPCR experiment, including sample preparation, fluorescent chemistries, primer/probe design, and data analysis applicable to cardiovascular research.

Original languageEnglish
Title of host publicationCardiac Gene Expression: Methods and Protocols
Pages121-143
Number of pages23
Volume366
DOIs
Publication statusPublished - 19 Mar 2007
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume366
ISSN (Print)10643745

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Ashton, K. J., & Headrick, J. P. (2007). Quantitative (Real-Time) RT-PCR in cardiovascular research. In Cardiac Gene Expression: Methods and Protocols (Vol. 366, pp. 121-143). (Methods in Molecular Biology; Vol. 366). https://doi.org/10.1385/1-59745-030-8:121