Hi-Plex2: A simple and robust approach to targeted sequencing-based genetic screening

Fleur Hammet, Khalid Mahmood, Tu Nguyen-Dumont, Melissa C. Southey, Daniel D. Buchanan, Andrew Lonie, Katherine L. Nathanson, Fergus J. Couch, Bernard J. Pope, Daniel J. Park

Research output: Contribution to journalArticleResearchpeer-review

8 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)118-122
Number of pages5
Issue number3
Publication statusPublished - 1 Sept 2019


  • Genetic screening
  • Hi-Plex
  • Multiplex PCR
  • Targeted DNA sequencing
  • Variant detection

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