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Sequencing of mRNA from Whole Blood using Nanopore Sequencing
Published on: June 3, 2019
INC-Seq: accurate single molecule reads using nanopore sequencing
Chenhao Li1,2, Kern Rei Chng1, Esther Jia Hui Boey1
1Genome Institute of Singapore, Singapore, 138672, Singapore.
Background:
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.
Results:
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 %.
Conclusions:
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.

