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Updated: Jun 13, 2026

Rup (RNA-seq Usability Assessment Pipeline) - Quality Control for Bulk RNA-seq Experiments in Eukaryotes
Published on: November 7, 2025
Quality assessment of transcriptome data using intrinsic statistical properties
Guillaume Brysbaert1, François-Xavier Pellay, Sebastian Noth
1Institut des Hautes Etudes Scientifiques & Institut de Recherche Interdisciplinaire (CNRS USR3078, Université de Lille1), 91440 Bures-sur-Yvette, France.
Abstract:
In view of potential application to biomedical diagnosis, tight transcriptome data quality control is compulsory. Usually, quality control is achieved using labeling and hybridization controls added at different stages throughout the processing of the biologic RNA samples. These control measures, however, only reflect the performance of the individual technical manipulations during the entire process and have no bearing as to the continued integrity of the RNA sample itself. Here we demonstrate that intrinsic statistical properties of the resulting transcriptome data signal and signal-variance distributions and their invariance can be identified independently of the animal species studied and the labeling protocol used. From these invariant properties we have developed a data model, the parameters of which can be estimated from individual experiments and used to compute relative quality measures based on similarity with large reference datasets. These quality measures add supplementary, non-redundant information to standard quality control estimates based on spike-in and hybridization controls, and are exploitable in data analysis. A software application for analyzing datasets as well as a reference dataset for AB1700 arrays are provided. They should allow AB1700 users to easily integrate this method into their analysis pipeline, and might instigate similar developments for other transcriptome platforms.
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