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.
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Ashton, K.J., Headrick, J.P. (2007). Quantitative (Real-Time) RT-PCR in Cardiovascular Research. In: Zhang, J., Rokosh, G. (eds) Cardiac Gene Expression. Methods in Molecular Biology, vol 366. Humana Press. https://doi.org/10.1007/978-1-59745-030-0_7
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DOI: https://doi.org/10.1007/978-1-59745-030-0_7
Publisher Name: Humana Press
Print ISBN: 978-1-58829-352-7
Online ISBN: 978-1-59745-030-0
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