期刊
MOLECULAR ECOLOGY RESOURCES
卷 14, 期 6, 页码 1097-1102出版社
WILEY-BLACKWELL
DOI: 10.1111/1755-0998.12324
关键词
bioinformatics; genomics; nanopore sequencing; next-generation sequencing
Oxford Nanopore's third-generation single-molecule sequencing platform promises to decrease costs for reagents and instrumentation. After a 2-year hiatus following the initial announcement, the first devices have been released as part of an early access program. We explore the performance of this platform by resequencing the lambda phage genome, and amplicons from a snake venom gland transcriptome. Although the handheld MinION sequencer can generate more than 150 megabases of raw data in one run, at most a quarter of the resulting reads map to the reference, with less than average 10% identity. Much of the sequence consists of insertion/deletion errors, or is seemingly without similarity to the template. Using the lambda phage data as an example, although the reads are long, averaging 5kb, at best 890 +/- 1932 bases per mapped read could be matched to the reference without soft clipping. In the course of a 36h run on the MinION, it was possible to resequence the 48kb lambda phage reference at 16x coverage. Currently, substantially larger projects would not be feasible using the MinION. Without increases in accuracy, which would be required for applications such as genome scaffolding and phasing, the current utility of the MinION appears limited. Library preparation requires access to a molecular laboratory, and is of similar complexity and cost to that of other next-generation sequencing platforms. The MinION is an exciting step in a new direction for single-molecule sequencing, though it will require dramatic decreases in error rates before it lives up to its promise.
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