Abridged adapter primers increase the target scope of Hi-Plex

Tú Nguyen-Dumont, Fleur Hammet, Maryam Mahmoodi, Bernard J. Pope, Graham G. Giles, John L. Hopper, Melissa C. Southey, Daniel J. Park

Research output: Contribution to journalArticleOtherpeer-review

7 Citations (Scopus)


Previously, we reported Hi-Plex, an amplicon-based method for targeted massively parallel sequencing capable of generating 60 amplicons simultaneously. In further experiments, however, we found our approach did not scale to higher amplicon numbers. Here, we report a modification to the original Hi-Plex protocol that includes the use of abridged adapter oligonucleotides as universal primers (bridge primers) in the initial PCR mixture. Full-length adapter primers (indexing primers) are included only during latter stages of thermal cycling with concomitant application of elevated annealing temperatures. Using this approach, we demonstrate the application of Hi-Plex across a broad range of amplicon numbers (16-plex, 62-plex, 250-plex, and 1003-plex) while preserving the low amount (25 ng) of input DNA required.

Original languageEnglish
Pages (from-to)33-36
Number of pages4
Issue number1
Publication statusPublished - 1 Jan 2015
Externally publishedYes


  • Genetic screening
  • Hi-Plex
  • Massively parallel sequencing
  • Mutation screening
  • Next-generation sequencing
  • Targeted sequencing

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