The effect of background noise and its removal on the analysis of single-cell expression data.

ABSTRACT:

Background

In droplet-based single-cell and single-nucleus RNA-seq experiments, not all reads associated with one cell barcode originate from the encapsulated cell. Such background noise is attributed to spillage from cell-free ambient RNA or barcode swapping events.

Results

Here, we characterize this background noise exemplified by three scRNA-seq and two snRNA-seq replicates of mouse kidneys. For each experiment, cells from two mouse subspecies are pooled, allowing to identify cross-genotype contaminating molecules and thus profile background noise. Background noise is highly variable across replicates and cells, making up on average 3-35% of the total counts (UMIs) per cell and we find that noise levels are directly proportional to the specificity and detectability of mar

SUBMITTER: Janssen P 

PROVIDER: S-EPMC10278251 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature