Dye spectra, dynamics and consequences

Roderic Fuerst IT-IS Life Science, Ireland

Abstract
The process of qPCR requires robust and reliable florescence monitoring of multiple dyes. Commonly qPCR systems use discrete excitation and emission filters in the monitoring of amplification and hybridization florescence. Also, algorithms used for resolving components of multiplex assays rely on the assumption that dye spectra remain constant during PCR and subsequent melting point analysis. In some cases this assumption is not valid. The nature and consequences of different approaches will be discussed.