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Abstract:
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As a common experimental technique, qPCR (Quantitative Real-time Polymerase Chain Reaction) is widely used to measure levels of nucleic acids, e.g., microRNAs in liquid biopsy. In addition to accurate and complete data, researchers have inevitably encountered uncertainly determined qPCR data because of intrinsically low overall amounts of biological materials and/or hidden contamination during sample collection or preparation. A lack of a reliable tool to account for the resulting statistical challenges may hinder biomarker detection, for example, identification and validation of microRNA biomarkers in liquid biopsies. We proposed and implemented a generalized cycle-to-threshold method (GCTOT) in a Web application, which can help identify contaminated qPCR data and account for low levels of biological materials in group comparisons. The R Shiny application provides user-friendly interfaces for researchers to easily input data and baseline information for analysis. The users can save statistical results and plots in their preferred formats. Our application shows that GCTOT may help identify interesting differential changes for biomarker detection and endpoint validation.
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