scnanoseq: an nf-core pipeline for Oxford Nanopore single-cell RNA-sequencing

Abstract

Recent advancements in long-read single-cell RNA sequencing (scRNA-seq) have facilitated the quantification of full-length transcripts and isoforms at the single-cell level. Historically, long-read data would need to be complemented with short-read single-cell data in order to overcome the higher sequencing errors to correctly identify cellular barcodes and unique molecular identifiers. Improvements in Oxford Nanopore sequencing, and development of novel computational methods have removed this requirement. Though these methods now exist, the limited availability of modular and portable workflows remains a challenge. Here we present, nf-core/scnanoseq, a secondary analysis pipeline for long-read single-cell and single-nuclei RNA that delivers gene and transcript-level quantification. The scnanoseq pipeline is implemented using Nextflow and is built upon the nf-core framework, enabling portability across computational environments, scalability and reproducibility of results across pipeline runs. The nf-core/scnanoseq workflow follows best practices for analyzing single-cell and single-nuclei data, performing barcode detection and correction, genome and transcriptome read alignment, unique molecular identifier deduplication, gene and transcript quantification, and extensive quality control reporting. AVAILABILITY OF DATA AND MATERIALSnf-core/scnanoseq is available at https://github.com/nf-core/scnanoseq under the MIT License and the documentation is available at https://nf-co.re/scnanoseq. The downstream analytical code for validation analysis is available at https://github.com/U-BDS/scnanoseq_analysis and all dataset sources have been disclosed under the methods section for each respective dataset.