Abstract
Analysis of single-cell RNA-seq data begins with pre-processing of sequencing reads to generate count matrices. We investigate algorithm choices for the challenges of pre-processing, and describe a workflow that balances efficiency and accuracy. Our workflow is based on the kallisto (https://pachterlab.github.io/kallisto/) and bustools (https://bustools.github.io/) programs, and is near-optimal in speed and memory. The workflow is modular, and we demonstrate its flexibility by showing how it can be used for RNA velocity analyses. Documentation and tutorials for using the kallisto | bus workflow are available at https://www.kallistobus.tools/.
Footnotes
Additional RNA velocity analysis (spliced vs standard workflow). Additional plot (MA plot) to benchmark panel figures. Additional info on IO in supplementary table. Additional references. Fixed typos. Updated all files.
