从福尔马林固定石蜡包埋的组织DNA中生成外显子组富集测序文库,用于下一代测序

Beth A. Marosy, Brian D. Craig, Kurt N. Hetrick, P. Dane Witmer, Hua Ling, Sean M. Griffith, Benjamin Myers, Elaine A. Ostrander, Janet L. Stanford, Lawrence C. Brody, Kimberly F. Doheny
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引用次数: 5

摘要

本单元描述了一种利用从福尔马林固定石蜡包埋(FFPE)样品中提取的DNA生成富集外显子组测序文库的技术。利用市售试剂盒,我们提出了一个低输入FFPE工作流程,从50 ng DNA开始。该程序包括一个修复步骤,以解决FFPE保存造成的损坏,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下一代短读测序平台进行测序。©2017 by John Wiley &儿子,Inc。
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Generating Exome Enriched Sequencing Libraries from Formalin-Fixed, Paraffin-Embedded Tissue DNA for Next-Generation Sequencing

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.

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Current Protocols in Human Genetics
Current Protocols in Human Genetics Biochemistry, Genetics and Molecular Biology-Genetics
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期刊介绍: Current Protocols in Human Genetics is the resource for designing and running successful research projects in all branches of human genetics.
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