title: The minimal amount of starting DNA for Agilent's hybrid capture-based targeted massively parallel sequencing.
abstract: Targeted capture massively parallel sequencing is increasingly being used in clinical settings, and as costs continue to decline, use of this technology may become routine in health care. However, a limited amount of tissue has often been a challenge in meeting quality requirements. To offer a practical guideline for the minimum amount of input DNA for targeted sequencing, we optimized and evaluated the performance of targeted sequencing depending on the input DNA amount. First, using various amounts of input DNA, we compared commercially available library construction kits and selected Agilent's SureSelect-XT and KAPA Biosystems' Hyper Prep kits as the kits most compatible with targeted deep sequencing using Agilent's SureSelect custom capture. Then, we optimized the adapter ligation conditions of the Hyper Prep kit to improve library construction efficiency and adapted multiplexed hybrid selection to reduce the cost of sequencing. In this study, we systematically evaluated the performance of the optimized protocol depending on the amount of input DNA, ranging from 6.25 to 200 ng, suggesting the minimal input DNA amounts based on coverage depths required for specific applications.
authors.affiliation: Samsung Biomedical Research Institute, Samsung Advanced Institute of Technology, Samsung Electronics Co. Ltd., Seoul 06351, Korea.|Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.|Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon 16419, Korea.
authors.forename: Jongsuk
authors.initials: J
authors.lastname: Chung
authors.affiliation: Samsung Biomedical Research Institute, Samsung Advanced Institute of Technology, Samsung Electronics Co. Ltd., Seoul 06351, Korea.|Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.
authors.forename: Dae-Soon
authors.initials: DS
authors.lastname: Son
authors.affiliation: Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.
authors.forename: Hyo-Jeong
authors.initials: HJ
authors.lastname: Jeon
authors.affiliation: Department of Pathology and Translational Genomics, Sungkyunkwan University School of Medicine, Suwon 16419, Korea.
authors.forename: Kyoung-Mee
authors.initials: KM
authors.lastname: Kim
authors.affiliation: Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.
authors.forename: Gahee
authors.initials: G
authors.lastname: Park
authors.affiliation: Office of Research and Development Strategy &Planning, Samsung Medical Center, Seoul 06351, Korea.
authors.forename: Gyu Ha
authors.initials: GH
authors.lastname: Ryu
authors.affiliation: Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.|Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon 16419, Korea.
authors.forename: Woong-Yang
authors.initials: WY
authors.lastname: Park
authors.affiliation: Samsung Biomedical Research Institute, Samsung Advanced Institute of Technology, Samsung Electronics Co. Ltd., Seoul 06351, Korea.|Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.
authors.forename: Donghyun
authors.initials: D
authors.lastname: Park
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D011933:Reagent Kits, Diagnostic; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods
chemical_list: D011933:Reagent Kits, Diagnostic; D004247:DNA