Empty drops in scRNA-seq uncover the surprising prevalence of sequestered neuropeptide mRNA and pervasive sequencing artifacts

Abstract The empty drops in single-cell sequencing experiments are an underexplored resource. As such, they present a substrate to ask questions orthogonal to standard single-cell sequencing workflows, calibrate statistical models using simple internal controls, and detect technical outliers which would be otherwise challenging to distinguish from real biology. In this case study, we report a relatively simple procedure to detect sequencing artifacts and make recommendations to reduce the risk of erroneous quantifications. In addition, we report the surprising abundance and co-expression of mRNA coding for neuropeptide-related genes in the empty drops, possibly reflecting underlying physiology.