Abstract:
:Reverse 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.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Freeman WM,Walker SJ,Vrana KEdoi
10.2144/99261rv01subject
Has Abstractpub_date
1999-01-01 00:00:00pages
112-22, 124-5issue
1eissn
0736-6205issn
1940-9818journal_volume
26pub_type
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