Internal oligo(dT) priming introduces systematic bias in bulk and single-cell RNA sequencing count data.

ABSTRACT: Significant advances in RNA sequencing have been recently made possible by using oligo(dT) primers for simultaneous mRNA enrichment and reverse transcription priming. The associated increase in efficiency has enabled more economical bulk RNA sequencing methods and the advent of high-throughput single-cell RNA sequencing, already one of the most widely adopted methods in transcriptomics. However, the effects of off-target oligo(dT) priming on gene expression quantification have not been appreciated. In the present study, we describe the extent, the possible causes, and the consequences of internal oligo(dT) priming across multiple public datasets obtained from various bulk and single-cell RNA sequencing platforms. To explore and address this issue, we developed a computational algorithm for

SUBMITTER: Svoboda M 

PROVIDER: S-EPMC9142200 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature