Ip Camilla L C, Loose Matthew, Tyson John R, de Cesare Mariateresa, Brown Bonnie L, Jain Miten, Leggett Richard M, Eccles David A, Zalunin Vadim, Urban John M, Piazza Paolo, Bowden Rory J, Paten Benedict, Mwaigwisya Solomon, Batty Elizabeth M, Simpson Jared T, Snutch Terrance P, Birney Ewan, Buck David, Goodwin Sara, Jansen Hans J, O'Grady Justin, Olsen Hugh E
Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
School of Life Sciences, Queens Medical Centre, University of Nottingham, Nottingham, UK.
F1000Res. 2015 Oct 15;4:1075. doi: 10.12688/f1000research.7201.1. eCollection 2015.
The advent of a miniaturized DNA sequencing device with a high-throughput contextual sequencing capability embodies the next generation of large scale sequencing tools. The MinION™ Access Programme (MAP) was initiated by Oxford Nanopore Technologies™ in April 2014, giving public access to their USB-attached miniature sequencing device. The MinION Analysis and Reference Consortium (MARC) was formed by a subset of MAP participants, with the aim of evaluating and providing standard protocols and reference data to the community. Envisaged as a multi-phased project, this study provides the global community with the Phase 1 data from MARC, where the reproducibility of the performance of the MinION was evaluated at multiple sites. Five laboratories on two continents generated data using a control strain of Escherichia coli K-12, preparing and sequencing samples according to a revised ONT protocol. Here, we provide the details of the protocol used, along with a preliminary analysis of the characteristics of typical runs including the consistency, rate, volume and quality of data produced. Further analysis of the Phase 1 data presented here, and additional experiments in Phase 2 of E. coli from MARC are already underway to identify ways to improve and enhance MinION performance.
一种具有高通量上下文测序能力的小型化DNA测序设备的出现，代表了下一代大规模测序工具。MinION™访问计划（MAP）于2014年4月由牛津纳米孔技术公司发起，使公众能够使用其通过USB连接的微型测序设备。MinION分析与参考联盟（MARC）由MAP的一部分参与者组成，旨在评估并为社区提供标准协议和参考数据。本研究作为一个多阶段项目，向全球社区提供了来自MARC的第一阶段数据，在多个地点评估了MinION性能的可重复性。来自两大洲的五个实验室使用大肠杆菌K-12的对照菌株生成数据，并根据修订后的ONT协议制备和测序样本。在此，我们提供了所使用协议的详细信息，以及对典型运行特征的初步分析，包括所产生数据的一致性、速率、量和质量。对这里呈现的第一阶段数据的进一步分析，以及MARC大肠杆菌第二阶段的其他实验已经在进行中，以确定改进和提高MinION性能的方法。
