A simplified hybrid capture approach retains high specificity and enables PCR-free workflow

We present a targeted sequencing workflow that eliminates bead-based capture, multiple washes, and post-hybridization PCR, while improving various aspects of data quality. The performance of the approach was evaluated by sequencing hundreds of samples and demonstrating high on-target rates, reduced duplicates, and improved indel accuracy. By combining the approach with a PCR-free library preparation, we enable an entirely PCR-free targeted sequencing assay which further improves indel calling, and shows the ability to call an HTT expansion, associated with Huntington's disease. This streamlined approach addresses key operational challenges in targeted sequencing, offering potential benefits for applications requiring rapid turnaround times or increased capability in variant detection.

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Publication Details

Journal
BMC Genomics
Published
2025-09-02
DOI
https://doi.org/10.1186/s12864-025-11939-6
Citations
4
Primary Topic
Molecular Biology Techniques and Applications
Type
article
Field-Weighted Citation Impact
3.22
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article

A simplified hybrid capture approach retains high specificity and enables PCR-free workflow

Kevin Green, Semyon Kruglyak, Bryan R. Lajoie, Joshua Chan et al.
4 citations
BMC Genomics
Molecular Biology Techniques and Applications
3.22
article

A simplified hybrid capture approach retains high specificity and enables PCR-free workflow

Kevin Green, Semyon Kruglyak, Bryan R. Lajoie, Joshua Chan, Xiaodong Qi, Michael J. R. Previte, Ramkrishna Adhikary, Peter T. Mains, Micah Ojeda, Andrew M. Boddicker, Laure Edoli, Adeline Mah, Kyle Metcalfe, Juan Moreno, Susan E. Cameron Devitt, Kelly Wiseman, Sophie Billings, Ben Krajacich, Junhua Zhao, Shawn Levy, Marina McCowin, Kyle Donohoe
article en
4 citations

Abstract

We present a targeted sequencing workflow that eliminates bead-based capture, multiple washes, and post-hybridization PCR, while improving various aspects of data quality. The performance of the approach was evaluated by sequencing hundreds of samples and demonstrating high on-target rates, reduced duplicates, and improved indel accuracy. By combining the approach with a PCR-free library preparation, we enable an entirely PCR-free targeted sequencing assay which further improves indel calling, and shows the ability to call an HTT expansion, associated with Huntington's disease. This streamlined approach addresses key operational challenges in targeted sequencing, offering potential benefits for applications requiring rapid turnaround times or increased capability in variant detection.

BMC GenomicsVol. 26(1)
Elements Biosciences (United States)
Openalex Percentile: Top 8%
Molecular Biology Techniques and Applications
3.22
DOIOpen Access PDFOpenAlex
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A simplified hybrid capture approach retains high specificity and enables PCR-free workflow — Kevin Green, Semyon Kruglyak, et al. · BMC Genomics (2025) | TGRS Research Map | TGRS