Journal article

Improved next-generation sequencing pre-capture library yields and sequencing parameters using on-bead PCR

Christopher R McEvoy, Timothy Semple, Bhargavi Yellapu, David Y Choong, Huiling Xu, Gisela Mir Arnau, Andrew P Fellowes, Stephen B Fox

BIOTECHNIQUES | FUTURE SCI LTD | Published : 2020

Abstract

Tumor DNA sequencing results can have important clinical implications. However, its use is often limited by low DNA input, owing to small tumor biopsy size. To help overcome this limitation we have developed a simple improvement to a commonly used next-generation sequencing (NGS) capture-based library preparation method using formalin-fixed paraffin-embedded-derived tumor DNA. By using on-bead PCR for pre-capture library generation we show that library yields are dramatically increased, resulting in decreased sample failure rates. Improved yields allowed for a reduction in PCR cycles, which translated into improved sequencing parameters without affecting variant calling. This methodology sho..

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