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Article
Quantitative (Real-Time) RT-PCR in cardiovascular research
Methods in Molecular Biology (2007)
  • Kevin John Ashton, Bond University
  • John Patrick Headrick
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.
Keywords
  • real-time quantitative polymerase chain reaction,
  • reverse transcription,
  • SYBR green I,
  • 5′-nuclease probes,
  • TaqMan,
  • molecular beacons,
  • standard curve,
  • comparative delta C T,
  • primer design
Disciplines
Publication Date
January 1, 2007
Publisher Statement
Citation only.

Kevin John Ashton and John Patrick Headrick. (2007) Quantitative (Real-Time) RT-PCR in cardiovascular research. Methods in Molecular Biology, 366(1), 121-143

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© Copyright Humana Press, 2007
Citation Information
Kevin John Ashton and John Patrick Headrick. "Quantitative (Real-Time) RT-PCR in cardiovascular research" Methods in Molecular Biology Vol. 366 Iss. 1 (2007)
Available at: http://works.bepress.com/kevin_ashton/1/