Cross-platform compatibility of Hi-Plex, a streamlined approach for targeted massively parallel sequencing.
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.
Genetic Epidemiology Laboratory, Department of Pathology, The University of Melbourne, Melbourne, Victoria 3010, Australia.
Tú
T
Nguyen-Dumont
Bernard J
BJ
Pope
Fleur
F
Hammet
Maryam
M
Mahmoodi
Helen
H
Tsimiklis
Melissa C
MC
Southey
Daniel J
DJ
Park
D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000191:economics / Q000379:methods*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000191:economics / Q000379:methods*; D013997:Time Factors
Disease gene screening; High-Plex PCR; Massively parallel sequencing; Molecular diagnostics; Targeted sequencing