System-specific periodicity in quantitative real-time polymerase chain reaction data questions threshold-based quantitation.

ABSTRACT: Real-time quantitative polymerase chain reaction (qPCR) data are found to display periodic patterns in the fluorescence intensity as a function of sample number for fixed cycle number. This behavior is seen for technical replicate datasets recorded on several different commercial instruments; it occurs in the baseline region and typically increases with increasing cycle number in the growth and plateau regions. Autocorrelation analysis reveals periodicities of 12 for 96-well systems and 24 for a 384-well system, indicating a correlation with block architecture. Passive dye experiments show that the effect may be from optical detector bias. Importantly, the signal periodicity manifests as periodicity in quantification cycle (C_q) values when these are estimated by the widely appli

SUBMITTER: Spiess AN 

PROVIDER: S-EPMC5154181 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature