评估牛津纳米孔技术MinION的性能

Q1 Biochemistry, Genetics and Molecular Biology Biomolecular Detection and Quantification Pub Date : 2015-03-01 DOI:10.1016/j.bdq.2015.02.001
T. Laver , J. Harrison , P.A. O’Neill , K. Moore , A. Farbos , K. Paszkiewicz , D.J. Studholme
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引用次数: 450

摘要

牛津纳米孔技术公司(ONT) MinION是一种新的测序技术,它可能提供数十千碱基(kb)的读取长度,仅受呈现给它的DNA分子长度的限制。该设备成本低,是目前可用的最便携的DNA测序仪,并且可以实时产生数据。它有许多潜在的应用,包括改善基因组序列组装和重复丰富区域的分辨率。在广泛采用这种技术之前,重要的是评估其性能和吞吐量和准确性方面的限制。在这项研究中,我们通过对三种不同核苷酸组成(从28.6%到70.7%)的细菌基因组进行重测序来评估MinION的性能;高G + C菌株在测序读数中代表性不足。我们估计MinION(基数调用后)的错误率为38.2%。平均和中位数读取长度分别为2 kb和1 kb,最长的单次读取长度为98 kb。一个5kb的rRNA操纵子全长被一个read覆盖。作为研究人员可用的第一个基于纳米孔的单分子测序仪,MinION是一个令人兴奋的前景;然而,目前的错误率限制了其与现有测序技术竞争的能力,尽管我们确实表明,MinION序列读取与Illumina MiSeq数据结合使用时,可以提高从头组装的连续性。
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Assessing the performance of the Oxford Nanopore Technologies MinION

The Oxford Nanopore Technologies (ONT) MinION is a new sequencing technology that potentially offers read lengths of tens of kilobases (kb) limited only by the length of DNA molecules presented to it. The device has a low capital cost, is by far the most portable DNA sequencer available, and can produce data in real-time. It has numerous prospective applications including improving genome sequence assemblies and resolution of repeat-rich regions. Before such a technology is widely adopted, it is important to assess its performance and limitations in respect of throughput and accuracy. In this study we assessed the performance of the MinION by re-sequencing three bacterial genomes, with very different nucleotide compositions ranging from 28.6% to 70.7%; the high G + C strain was underrepresented in the sequencing reads. We estimate the error rate of the MinION (after base calling) to be 38.2%. Mean and median read lengths were 2 kb and 1 kb respectively, while the longest single read was 98 kb. The whole length of a 5 kb rRNA operon was covered by a single read. As the first nanopore-based single molecule sequencer available to researchers, the MinION is an exciting prospect; however, the current error rate limits its ability to compete with existing sequencing technologies, though we do show that MinION sequence reads can enhance contiguity of de novo assembly when used in conjunction with Illumina MiSeq data.

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来源期刊
Biomolecular Detection and Quantification
Biomolecular Detection and Quantification Biochemistry, Genetics and Molecular Biology-Biochemistry
CiteScore
14.20
自引率
0.00%
发文量
0
审稿时长
8 weeks
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