Genome Institute of Singapore, Singapore, 138672, Singapore.
Department of Computer Science, National University of Singapore, Singapore, 117417, Singapore.
Gigascience. 2016 Aug 2;5(1):34. doi: 10.1186/s13742-016-0140-7.
Nanopore sequencing provides a rapid, cheap and portable real-time sequencing platform with the potential to revolutionize genomics. However, several applications are limited by relatively high single-read error rates (>10 %), including RNA-seq, haplotype sequencing and 16S sequencing.
We developed the Intramolecular-ligated Nanopore Consensus Sequencing (INC-Seq) as a strategy for obtaining long and accurate nanopore reads, starting with low input DNA. Applying INC-Seq for 16S rRNA-based bacterial profiling generated full-length amplicon sequences with a median accuracy >97 %.
INC-Seq reads enabled accurate species-level classification, identification of species at 0.1 % abundance and robust quantification of relative abundances, providing a cheap and effective approach for pathogen detection and microbiome profiling on the MinION system.
纳米孔测序提供了一种快速、廉价且便携的实时测序平台，具有彻底改变基因组学的潜力。然而，一些应用受到相对较高的单读错误率（>10%）的限制，包括 RNA-seq、单倍型测序和 16S 测序。
我们开发了分子内连接的纳米孔一致测序（INC-Seq）作为一种策略，从低输入 DNA 开始获得长而准确的纳米孔读数。应用 INC-Seq 进行基于 16S rRNA 的细菌分析，生成了具有中位数准确率 >97%的全长扩增子序列。
INC-Seq 读数可实现准确的物种分类，鉴定丰度为 0.1%的物种，并对相对丰度进行稳健定量，为 MinION 系统上的病原体检测和微生物组分析提供了一种廉价有效的方法。
