纳米孔测序
单细胞测序
计算生物学
条形码
RNA剪接
计算机科学
基因组学
选择性拼接
单细胞分析
DNA测序
吞吐量
深度测序
转录组
生物
Illumina染料测序
顺序装配
细胞生物学
大规模并行测序
基因组
外显子组测序
核糖核酸
遗传学
基因
基因亚型
细胞
表型
操作系统
电信
无线
作者
Kevin Lebrigand,Virginie Magnone,Pascal Barbry,Rainer Waldmann
标识
DOI:10.1038/s41467-020-17800-6
摘要
Abstract Droplet-based high throughput single cell sequencing techniques tremendously advanced our insight into cell-to-cell heterogeneity. However, those approaches only allow analysis of one extremity of the transcript after short read sequencing. In consequence, information on splicing and sequence heterogeneity is lost. To overcome this limitation, several approaches that use long-read sequencing were introduced recently. Yet, those techniques are limited by low sequencing depth and/or lacking or inaccurate assignment of unique molecular identifiers (UMIs), which are critical for elimination of PCR bias and artifacts. We introduce ScNaUmi-seq, an approach that combines the high throughput of Oxford Nanopore sequencing with an accurate cell barcode and UMI assignment strategy. UMI guided error correction allows to generate high accuracy full length sequence information with the 10x Genomics single cell isolation system at high sequencing depths. We analyzed transcript isoform diversity in embryonic mouse brain and show that ScNaUmi-seq allows defining splicing and SNVs (RNA editing) at a single cell level.
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