Genetic Epidemiology Laboratory, Department of Pathology, The University of Melbourne, Melbourne, Victoria 3010, Australia.
Anal Biochem. 2013 Nov 15;442(2):127-9. doi: 10.1016/j.ab.2013.07.046. Epub 2013 Aug 8.
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.
尽管近年来每碱基测序成本有所降低，但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复杂的方案的限制。为了解决这些限制，我们之前开发了 Hi-Plex，这是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，我们证明了使用杂交接头的 Hi-Plex 可以生成适合 Ion Torrent 和 TruSeq 化学的文库，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-Plex 的功能，使其成为各种遗传筛选应用的准确、经济、灵活和快速的方法。
