PeerJ | |
Effective ribosomal RNA depletion for single-cell total RNA-seq by scDASH | |
article | |
Danson S.C. Loi1  Lei Yu1  Angela R. Wu1  | |
[1] Division of Life Science, Hong Kong University of Science and Technology;Department of Chemical and Biological Engineering, Hong Kong University of Science and Technology;Hong Kong Branch of Guangdong Southern Marine Science and Engineering Laboratory ,(Guangzhou), Hong Kong University of Science and Technology | |
关键词: Single-cell transcriptomics; scRNA-seq; rRNA depletion; CRISPR; | |
DOI : 10.7717/peerj.10717 | |
学科分类:社会科学、人文和艺术(综合) | |
来源: Inra | |
【 摘 要 】
A decade since its invention, single-cell RNA sequencing (scRNA-seq) has become a mainstay technology for profiling transcriptional heterogeneity in individual cells. Yet, most existing scRNA-seq methods capture only polyadenylated mRNA to avoid the cost of sequencing non-messenger transcripts, such as ribosomal RNA (rRNA), that are usually not of-interest. Hence, there are not very many protocols that enable single-cell analysis of total RNA. We adapted a method called DASH (Depletion of Abundant Sequences by Hybridisation) to make it suitable for depleting rRNA sequences from single-cell total RNA-seq libraries. Our analyses show that our single-cell DASH (scDASH) method can effectively deplete rRNAs from sequencing libraries with minimal off-target non-specificity. Importantly, as a result of depleting the rRNA, the rest of the transcriptome is significantly enriched for detection.
【 授权许可】
CC BY
【 预 览 】
Files | Size | Format | View |
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RO202307100006767ZK.pdf | 13133KB | download |