从福尔马林固定、石蜡包埋组织DNA中生成外显子富集测序文库用于下一代测序。
Generating Exome Enriched Sequencing Libraries from Formalin-Fixed, Paraffin-Embedded Tissue DNA for Next-Generation Sequencing.
作者信息
Marosy Beth A, Craig Brian D, Hetrick Kurt N, Witmer P Dane, Ling Hua, Griffith Sean M, Myers Benjamin, Ostrander Elaine A, Stanford Janet L, Brody Lawrence C, Doheny Kimberly F
机构信息
Center for Inherited Disease Research, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland.
National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland.
出版信息
Curr Protoc Hum Genet. 2017 Jan 11;92:18.10.1-18.10.25. doi: 10.1002/cphg.27.
Abstract
This unit describes a technique for generating exome-enriched sequencing libraries using DNA extracted from formalin-fixed paraffin-embedded (FFPE) samples. Utilizing commercially available kits, we present a low-input FFPE workflow starting with 50 ng of DNA. This procedure includes a repair step to address damage caused by FFPE preservation that improves sequence quality. Subsequently, libraries undergo an in-solution-targeted selection for exons, followed by sequencing using the Illumina next-generation short-read sequencing platform. © 2017 by John Wiley & Sons, Inc.
摘要
本单元描述了一种使用从福尔马林固定石蜡包埋（FFPE）样本中提取的DNA生成外显子富集测序文库的技术。利用市售试剂盒，我们展示了一种低输入量的FFPE工作流程，起始DNA量为50 ng。该程序包括一个修复步骤，以解决FFPE保存造成的损伤，从而提高序列质量。随后，文库进行溶液内靶向外显子选择，然后使用Illumina下一代短读长测序平台进行测序。© 2017约翰威立国际出版公司。
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