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Mostly natural sequencing-by-synthesis for scRNA-seq using Ultima sequencing
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Mostly natural sequencing-by-synthesis for scRNA-seq using Ultima sequencing
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Journal Article
Mostly natural sequencing-by-synthesis for scRNA-seq using Ultima sequencing
2023
Overview
Here we introduce a mostly natural sequencing-by-synthesis (mnSBS) method for single-cell RNA sequencing (scRNA-seq), adapted to the Ultima genomics platform, and systematically benchmark it against current scRNA-seq technology. mnSBS uses mostly natural, unmodified nucleotides and only a low fraction of fluorescently labeled nucleotides, which allows for high polymerase processivity and lower costs. We demonstrate successful application in four scRNA-seq case studies of different technical and biological types, including 5′ and 3′ scRNA-seq, human peripheral blood mononuclear cells from a single individual and in multiplex, as well as Perturb-Seq. Benchmarking shows that results from mnSBS-based scRNA-seq are very similar to those using Illumina sequencing, with minor differences in results related to the position of reads relative to annotated gene boundaries, owing to single-end reads of Ultima being closer to gene ends than reads from Illumina. The method is thus compatible with state-of-the-art scRNA-seq libraries independent of the sequencing technology. We expect mnSBS to be of particular utility for cost-effective large-scale scRNA-seq projects.
A mostly natural sequencing-by-synthesis method is effective for scRNA-seq, particularly in large-scale studies.
Publisher
Nature Publishing Group US,Nature Publishing Group
