Consistent ultra-long DNA sequencing with automated slow pipetting.
Consistent ultra-long DNA sequencing with automated slow pipetting.
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一致的超长DNA测序与自动缓慢移液。
DOI：
10.1186/s12864-021-07500-w
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发表时间：
2021-03-12
期刊：
影响因子：
4.4
通讯作者：
O'Connor DH
中科院分区：
文献类型：
--
作者：
Prall TM;Neumann EK;Karl JA;Shortreed CG;Baker DA;Bussan HE;Wiseman RW;O'Connor DH
Oxford Nanopore Technologies的仪器可以对很长的读数进行测序。长读段通过明确跨越基因组的重复元件来改善序列组装。显着长度的测序读数需要通过尽可能慢地移液试剂以最大限度地减少剪切来通过文库制备来保存长DNA模板分子。这个过程是耗时的，并且在保留读取长度方面是不一致的，因为即使体积流速的微小变化也会导致模板剪切。我们设计了SNAILS（Slow Nucleic Acid Instrument for Long Sequences），这是一种3D打印仪器，可自动缓慢移液用于牛津纳米孔测序的长读文库制备中的试剂。在六个测序文库中，SNAILS保留了长度超过100个碱基的更多读段，并在手动缓慢移液过程中增加了其文库的平均读段长度。SNAILS是一种低成本、易于部署的解决方案，用于改进需要显着长度读取的测序项目。通过自动化文库制备试剂的缓慢移液，SNAILS提高了长读段纳米孔测序的一致性和通量。在线版本包含补充材料，可通过10.1186/s12864-021-07500-w获取。
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. 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. 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. The online version contains supplementary material available at 10.1186/s12864-021-07500-w.
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