title: Library construction for next-generation sequencing: overviews and challenges.
abstract: High-throughput sequencing, also known as next-generation sequencing (NGS), has revolutionized genomic research. In recent years, NGS technology has steadily improved, with costs dropping and the number and range of sequencing applications increasing exponentially. Here, we examine the critical role of sequencing library quality and consider important challenges when preparing NGS libraries from DNA and RNA sources. Factors such as the quantity and physical characteristics of the RNA or DNA source material as well as the desired application (i.e., genome sequencing, targeted sequencing, RNA-seq, ChIP-seq, RIP-seq, and methylation) are addressed in the context of preparing high quality sequencing libraries. In addition, the current methods for preparing NGS libraries from single cells are also discussed.
authors: NGS and Microarray Core Facility, The Scripps Research Institute, La Jolla, CA.
authors: Steven R
authors: SR
authors: Head
authors: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA.
authors: H Kiyomi
authors: HK
authors: Komori
authors: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA.
authors: Sarah A
authors: SA
authors: LaMere
authors: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA.
authors: Thomas
authors: T
authors: Whisenant
authors: Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium.
authors: Filip
authors: F
authors: Van Nieuwerburgh
authors: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA.
authors: Daniel R
authors: DR
authors: Salomon
authors: NGS and Microarray Core Facility, The Scripps Research Institute, La Jolla, CA.
authors: Phillip
authors: P
authors: Ordoukhanian
mesh_terms: D000818:Animals; D001709:Biotechnology; D019175:DNA Methylation; D015723:Gene Library; D023281:Genomics; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017423:Sequence Analysis, RNA
keywords: ChIP-seq; DNA; DNA-seq; RIP-seq; RNA; RNA-seq; deep sequencing; library preparation; next-generation sequencing