Hi-Plex2: a simple and robust approach to targeted sequencing-based genetic screening
Hi-Plex2: a simple and robust approach to targeted sequencing-based genetic screening
复制标题
DOI：
10.2144/btn-2019-0026
复制
发表时间：
2019-09-01
期刊：
影响因子：
2.7
通讯作者：
Park, Daniel J.
中科院分区：
文献类型：
--
作者：
Hammet, Fleur;Mahmood, Khalid;Park, Daniel J.
我们之前报道过Hi-Plex，这是一种用于构建靶向DNA测序文库的多重PCR方法，它提供了一种与高通量处理兼容的低成本协议。在这里，我们详细介绍了一种改进的协议Hi-Plex2，它可以更有效地构建中小型面板大小的库，同时保持Hi-Plex平台的低成本、简单性和准确性。Hi-Plex2应用于三个面板，包括291,740和1193个扩增子，靶向与乳腺癌和/或结肠癌风险相关的基因。我们展示了脱靶扩增的大幅减少，使得使用以前的Hi-Plex化学无法实现的中小型设计面板的库构建成为可能。hi - plex2是一种基于增强型PCR扩增子的方法，用于构建中小型靶向DNA测序文库，该方法结合了简单的改进以最大限度地减少脱靶效应，同时保留了其前身Hi-Plex的优点。
We have previously reported Hi-Plex, a multiplex PCR methodology for building targeted DNA sequencing libraries that offers a low-cost protocol compatible with high-throughput processing. Here, we detail an improved protocol, Hi-Plex2, that more effectively enables the robust construction of small-to-medium panel-size libraries while maintaining low cost, simplicity and accuracy benefits of the Hi-Plex platform. Hi-Plex2 was applied to three panels, comprising 291, 740 and 1193 amplicons, targeting genes associated with risk for breast and/or colon cancer. We show substantial reduction of off-target amplification to enable library construction for small-to-medium sized design panels not possible using the previous Hi-Plex chemistry.METHOD SUMMARYHi-Plex2 is an enhanced PCR amplicon-based method for the construction of small-to-medium-sized targeted DNA sequencing libraries that incorporates simple improvements to minimise off-target effects while retaining the benefits of its predecessor, Hi-Plex.