Cross-platform compatibility of Hi-Plex, a streamlined approach for targeted massively parallel sequencing.
Cross-platform compatibility of Hi-Plex, a streamlined approach for targeted massively parallel sequencing.
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Hi-Plex 的跨平台兼容性，是一种用于定向大规模并行测序的简化方法。
DOI：
10.1016/j.ab.2013.07.046
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发表时间：
2013
影响因子：
2.9
通讯作者：
Park,DanielJ
中科院分区：
文献类型：
--
作者：
Nguyen-Dumont,Tu;Pope,BernardJ;Hammet,Fleur;Mahmoodi,Maryam;Tsimiklis,Helen;Southey,MelissaC;Park,DanielJ
尽管近年来每碱基测序成本已经降低，但是用于靶向大规模平行测序的文库制备仍然受到高试剂成本、有限的设计灵活性和方案复杂性的限制。为了解决这些限制，我们以前开发了Hi-PCR，一种用于筛选基因组靶区域的聚合酶链反应（PCR）大规模平行测序策略。在这里，我们证明了与混合衔接子一起应用的Hi-Plex可以生成适合使用Ion Torrent和TruSeq化学物质进行测序的文库，并且调整引物浓度可以提高覆盖均匀性。这些结果扩展了Hi-PCR的能力，使其成为一种准确、经济、灵活和快速的方法，用于各种遗传筛查应用。
Although per-base sequencing costs have decreased during recent years, library preparation for targeted massively parallel sequencing remains constrained by high reagent cost, limited design flexibility, and protocol complexity. To address these limitations, we previously developed Hi-Plex, a polymerase chain reaction (PCR) massively parallel sequencing strategy for screening panels of genomic target regions. Here, we demonstrate that Hi-Plex applied with hybrid adapters can generate a library suitable for sequencing with both the Ion Torrent and the TruSeq chemistries and that adjusting primer concentrations improves coverage uniformity. These results expand Hi-Plex capabilities as an accurate, affordable, flexible, and rapid approach for various genetic screening applications.