System-Specific Periodicity in qPCR Data and its Impact on Quantitation
Poster May 14, 2015
Joel Tellinghuisen,1 Stefan Roediger,2 Thomas Volksdorf,3 and Andrej-Nikolai Spiess3
Statistical noise is a feature of every quantitative PCR (qPCR) curve. In principle, two different forms of noise can be encountered: (i) the dispersion of fluorescence signals about their true values cycle to cycle (within-sample noise) and (ii) the dispersion between different replicate qPCR curves at the same cycle (between-sample noise). The most obvious between-sample noise effect is an overall shifting of the qPCR curve on the y-axis, sometimes with scaling of the signal above baseline, such that both the baseline and the plateau-baseline difference may vary reaction to reaction. These effects have long been known, but not fully understood.
In recent work (Tellinghuisen & Spiess, 2014), we showed preliminary results on a published large-scale technical replicate dataset (Ruijter et al., 2013) that revealed unexpected between-sample periodicity for fluorescence values at all cycle numbers. The origin of these periodic patterns in qPCR data remains elusive.
To examine this phenomenon in more detail, we have employed autocorrelation analysis on a larger cohort of published and self-generated high-replicate qPCR data acquired from different platforms and have analysed Cq values with respect to intrinsic periodic patterns when obtained under several different quantitation regimes.
Analysis of the Effect of Aggregated β-Amyloid on Cellular Signaling Pathways Critical for Memory in Alzheimer’s DiseasePoster
Here we evaluate the ability to detect changes in phosphorylation levels of ERK and CREB following treatment with Aβ using the SH-SY5Y neuroblastoma cell line.READ MORE
Characterizing GPCR Activation Using Automated Live Cell ImagingPoster
Here we describe a live cell imaging based approach to detect GPCR activation using the Lionheart™ FX Automated Live Cell Imager and Gen5™ Microplate Reader and Imager Software.READ MORE
Assessment of Oral LISPRO Treatment in Ameliorating Amyloid and Tau Pathology in Transgenic Alzheimer’s Mice ModelPoster
Ionic co-crystals of lithium salicylate with organic proline (LISPRO) showed better safety and pharmacokinetic profile of lithium in plasma and brain of wild-type and transgenic Alzheimer mice model compared to lithium salts.READ MORE