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

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

doi:10.2144/btn-2019-0026

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

Fleur Hammet, Khalid Mahmood, Thomas R. Green, 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 journal › Article › peer-review

5 Scopus citations

Abstract

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 language	English (US)
Pages (from-to)	118-122
Number of pages	5
Journal	BioTechniques
Volume	67
Issue number	3
DOIs	https://doi.org/10.2144/btn-2019-0026
State	Published - 2019

Keywords

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

ASJC Scopus subject areas

Biotechnology
Biochemistry, Genetics and Molecular Biology(all)

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Cite this

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title = "Hi-Plex2: A simple and robust approach to targeted sequencing-based genetic screening",

abstract = "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.",

keywords = "Genetic screening, Hi-Plex, Multiplex PCR, Targeted DNA sequencing, Variant detection",

author = "Fleur Hammet and Khalid Mahmood and Green, {Thomas R.} and Tu Nguyen-Dumont and Southey, {Melissa C.} and Buchanan, {Daniel D.} and Andrew Lonie and Nathanson, {Katherine L.} and Couch, {Fergus J.} and Pope, {Bernard J.} and Park, {Daniel J.}",

note = "Publisher Copyright: {\textcopyright} 2019 Daniel Park.",

year = "2019",

doi = "10.2144/btn-2019-0026",

language = "English (US)",

volume = "67",

pages = "118--122",

journal = "BioTechniques",

issn = "0736-6205",

publisher = "Eaton Publishing Company",

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T2 - A simple and robust approach to targeted sequencing-based genetic screening

AU - Hammet, Fleur

AU - Mahmood, Khalid

AU - Green, Thomas R.

AU - Nguyen-Dumont, Tu

AU - Southey, Melissa C.

AU - Buchanan, Daniel D.

AU - Lonie, Andrew

AU - Nathanson, Katherine L.

AU - Couch, Fergus J.

AU - Pope, Bernard J.

AU - Park, Daniel J.

PY - 2019

Y1 - 2019

N2 - 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.

AB - 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.

KW - Genetic screening

KW - Hi-Plex

KW - Multiplex PCR

KW - Targeted DNA sequencing

KW - Variant detection

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IS - 3

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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