Novel, Fully Automated Method Allows Efficient Analysis of qPCR Data for Qualitative Calling Based on Comparative Cq
Poster Apr 30, 2012
Collaborative research by Pioneer Hi-Bred (a DuPont company) and Azure PCR Limited
qPCR is extensively applied to determine the identity of genetically modified (GM) seeds and plants. Analysis and interpretation of qPCR data for any application is limited by sample variability1, poor assay performance1 and arbitrarily set thresholds2 which can all lead to ambiguous and subjective result calling that is reliant upon the expertise and experience of the scientist interpreting the assay. Processing of raw data output from thermal cyclers using analysis software, to provide data more easily interpretable by the human eye, can lead to loss of data3 and error prone sample calling4,5. In this study we retrospectively examined data produced in qPCR-based plant genotyping tests at Pioneer Hi-Bred to assess if patent-pending AzurePCR automated analysis produced similar results without the need for manual input of analysis parameters or manipulation of raw data.
When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.READ MORE