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Modeling real-time PCR kinetics: Richards reparametrized equation for quantitative estimation of european hake (Merluccius merluccius)

AuthorsSánchez, Ana Cristina ; Vázquez, José Antonio ; Quinteiro, Javier; González Sotelo, Carmen
KeywordsReal-time PCR
Merluccius merluccius identification
DNA quantification
Mathematical modeling
Richards equation
Issue Date2013
PublisherAmerican Chemical Society
CitationJournal of Agricultural and Food Chemistry 61(14): 3488-3493 (2013)
AbstractReal-time PCR is the most sensitive method for detection and precise quantification of specific DNA sequences, but it is not usually applied as a quantitative method in seafood. In general, benchmark techniques, mainly cycle threshold (Ct), are the routine method for quantitative estimations, but they are not the most precise approaches for a standard assay. In the present work, amplification data from European hake (Merluccius merluccius) DNA samples were accurately modeled by three sigmoid reparametrized equations, where the lag phase parameter (λc) from the Richards equation with four parameters was demonstrated to be the perfect substitute for Ct for PCR quantification. The concentrations of primers and probes were subsequently optimized by means of that selected kinetic parameter. Finally, the linear correlation among DNA concentration and λc was also confirmed.
Description6 páginas, 4 figuras, 4 tablas
Publisher version (URL)http://dx.doi.org/10.1021/jf400136j
Appears in Collections:(IIM) Artículos
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