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pubmed-article:9894600pubmed:abstractTextReverse transcription PCR (RT-PCR) represents a sensitive and powerful tool for analyzing RNA. While it has tremendous potential for quantitative applications, a comprehensive knowledge of its technical aspects is required. Successful quantitative RT-PCR involves correction for experimental variations in individual RT and PCR efficiencies. This review addresses the mathematics of RT-PCR, choice of RNA standards (internal vs. external) and quantification strategies (competitive, noncompetitive and kinetic [real-time] amplification). Finally, the discussion turns to practical considerations in experimental design. It is hoped that this review will be appropriate for those undertaking these experiments for the first time or wishing to improve (or validate) a technique in what is frequently a confusing and contradictory field.lld:pubmed
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pubmed-article:9894600pubmed:authorpubmed-author:FreemanW MWMlld:pubmed
pubmed-article:9894600pubmed:authorpubmed-author:WalkerS JSJlld:pubmed
pubmed-article:9894600pubmed:authorpubmed-author:VranaK EKElld:pubmed
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pubmed-article:9894600pubmed:pagination112-22, 124-5lld:pubmed
pubmed-article:9894600pubmed:dateRevised2007-11-14lld:pubmed
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pubmed-article:9894600pubmed:articleTitleQuantitative RT-PCR: pitfalls and potential.lld:pubmed
pubmed-article:9894600pubmed:affiliationWake Forest University School of Medicine, Winston-Salem, NC, USA.lld:pubmed
pubmed-article:9894600pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:9894600pubmed:publicationTypeResearch Support, U.S. Gov't, P.H.S.lld:pubmed
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