A high-plex PCR approach for massively parallel sequencing

T Nguyen-Dumont; BJ Pope; F Hammet; MC Southey; DJ Park

Journal article

A high-plex PCR approach for massively parallel sequencing

T Nguyen-Dumont, BJ Pope, F Hammet, MC Southey, DJ Park

Biotechniques | FUTURE SCI LTD | Published : 2013

DOI: 10.2144/000114052

Abstract

Current methods for targeted massively parallel sequencing (MPS) have several drawbacks, including limited design flexibility, expense, and protocol complexity, which restrict their application to settings involving modest target size and requiring low cost and high throughput. To address this, we have developed Hi-Plex, a PCR-MPS strategy intended for high-throughput screening of multiple genomic target regions that integrates simple, automated primer design software to control product size. Featuring permissive thermocycling conditions and clamp bias reduction, our protocol is simple, cost- and time-effective, uses readily available reagents, does not require expensive instrumentation, and..

View full abstract

University of Melbourne Researchers

Tu Nguyen Author

Bernard Pope Author

Melissa Southey Author

Daniel Park Author

Related Projects (2)

Translation of information about the gene PALB2 into breast cancer clinical genetic services

Today in Australia women attending clinical genetics services and receiving genetic counselling due to a personal and/or family history of b..

Next generation DNA-based diagnostics and discovery

The next generation of medical diagnostics and discovery in disease research will involve the marriage of PCR, a tool used to amplify large ..

Grants

Awarded by Australian National Health and Medical Research Council (NHMRC)

Awarded by Victorian Life Sciences Computation Initiative (VLSCI)

Funding Acknowledgements

This work was supported by the Australian National Health and Medical Research Council (NHMRC) (APP1025879 and APP1029974), by a Victorian Life Sciences Computation Initiative (VLSCI) grant number VR0182 on its Peak Computing Facility at the University of Melbourne, an initiative of the Victorian Government, and by the Victorian Breast Cancer Research Consortium (VBCRC). TN-D is a Susan G. Komen for the Cure Postdoctoral Fellow. MCS is a VBCRC Group Leader and Senior Research Fellow of the NHMRC. We thank the Australian Breast Cancer Family Study (ABCFS, Principal Investigator John Hopper) for providing the cell-line derived DNA, and Ee Ming Wong for providing the FFPE tumor-derived DNA.