自动化慢速移液的一致超长 DNA 测序。
Consistent ultra-long DNA sequencing with automated slow pipetting.
机构信息
Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison, USA.
Wisconsin National Primate Research Center, University of Wisconsin, Madison, USA.
出版信息
BMC Genomics. 2021 Mar 12;22(1):182. doi: 10.1186/s12864-021-07500-w.
BACKGROUND
Oxford Nanopore Technologies' instruments can sequence reads of great length. Long reads improve sequence assemblies by unambiguously spanning repetitive elements of the genome. Sequencing reads of significant length requires the preservation of long DNA template molecules through library preparation by pipetting reagents as slowly as possible to minimize shearing. This process is time-consuming and inconsistent at preserving read length as even small changes in volumetric flow rate can result in template shearing.
RESULTS
We have designed SNAILS (Slow Nucleic Acid Instrument for Long Sequences), a 3D-printable instrument that automates slow pipetting of reagents used in long read library preparation for Oxford Nanopore sequencing. Across six sequencing libraries, SNAILS preserved more reads exceeding 100 kilobases in length and increased its libraries' average read length over manual slow pipetting.
CONCLUSIONS
SNAILS is a low-cost, easily deployable solution for improving sequencing projects that require reads of significant length. By automating the slow pipetting of library preparation reagents, SNAILS increases the consistency and throughput of long read Nanopore sequencing.
背景
牛津纳米孔技术公司的仪器可以对长度非常长的读段进行测序。长读段通过明确跨越基因组重复元件来提高序列组装质量。要获得显著长度的测序读段，需要通过尽可能缓慢地移液试剂来制备文库，以最小化剪切，从而保持长 DNA 模板分子。这个过程既耗时又不一致，因为即使体积流速的微小变化也可能导致模板剪切。
结果
我们设计了 SNAILS（用于长序列的缓慢核酸仪器），这是一种 3D 可打印仪器，可自动缓慢移液用于牛津纳米孔测序的长读段文库制备的试剂。在六个测序文库中，SNAILS 保留了更多长度超过 100 千碱基的读段，并增加了其文库的平均读段长度，超过了手动缓慢移液。
结论
SNAILS 是一种低成本、易于部署的解决方案，可用于提高需要显著长度读段的测序项目。通过自动缓慢移液文库制备试剂，SNAILS 提高了长读段纳米孔测序的一致性和通量。
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