NEBNext Direct: A Novel, Rapid, Hybridization-Based Approach for the Capture and Library Conversion of Genomic Regions of Interest
| Autor: | Noa Henig, Amy B. Emerman, Kruti M. Patel, Sarah K. Bowman, Andrew Barry, Andrew F. Gardner, Cynthia L. Hendrickson |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Genetics Time Factors Library preparation 0206 medical engineering High-Throughput Nucleotide Sequencing Nucleic Acid Hybridization 02 engineering and technology General Medicine Computational biology Genomics Biology Target enrichment DNA sequencing 03 medical and health sciences Identification (information) 030104 developmental biology Workflow Target capture Copy-number variation 020602 bioinformatics Gene Library |
| Zdroj: | Current Protocols in Molecular Biology |
| ISSN: | 1934-3647 |
| Popis: | Next-generation sequencing (NGS) is a powerful tool for genomic studies, translational research, and clinical diagnostics that enables the detection of single nucleotide polymorphisms, insertions and deletions, copy number variations, and other genetic variations. Target enrichment technologies improve the efficiency of NGS by only sequencing regions of interest, which reduces sequencing costs while increasing coverage of the selected targets. Here we present NEBNext Direct®, a hybridization-based, target-enrichment approach that addresses many of the shortcomings of traditional target-enrichment methods. This approach features a simple, 7-hr workflow that uses enzymatic removal of off-target sequences to achieve a high specificity for regions of interest. Additionally, unique molecular identifiers are incorporated for the identification and filtering of PCR duplicates. The same protocol can be used across a wide range of input amounts, input types, and panel sizes, enabling NEBNext Direct to be broadly applicable across a wide variety of research and diagnostic needs. © 2017 by John Wiley & Sons, Inc. Keywords: in-solution hybridization; Illumina library preparation; next-generation sequencing; NGS library preparation; NGS target capture; PCR enrichment; target enrichment |
| Databáze: | OpenAIRE |
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