Single-cell and single-nucleus RNA sequencing have been widely adopted in studies of heterogeneous tissues to estimate their cellular composition and obtain transcriptional profiles of individual cells. However, the current fragmentary understanding of artefacts introduced by sample preparation protocols impedes the selection of optimal workflows and compromises data interpretation. To bridge this gap, researchers at the University of Western Australia compared performance of several workflows applied to adult mouse kidneys. This study encompasses two tissue dissociation protocols, two cell preservation methods, bulk tissue RNA sequencing, single-cell and three single-nucleus RNA sequencing workflows for the 10x Genomics Chromium platform. These experiments enable a systematic comparison of recovered cell types and their transcriptional profiles across the workflows and highlight protocol-specific biases important for the experimental design and data interpretation.
Systematic bias assessment in solid tissue 10x scRNA-seq workflows
Denisenko E et al. (2019) Systematic bias assessment in solid tissue 10x scRNA-seq workflows. bioRXiv [Epub ahead of print]. [abstract]