从FFPE组织中提取DNA的一种经济有效且可扩展的方法。

IF 2.5 Q3 BIOCHEMICAL RESEARCH METHODS
Biology Methods and Protocols Pub Date : 2025-01-10 eCollection Date: 2025-01-01 DOI:10.1093/biomethods/bpaf003
Christoph Geisenberger, Edgar Chimal, Philipp Jurmeister, Frederick Klauschen
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引用次数: 0

摘要

癌症的基因组图谱在诊断和治疗计划中发挥着越来越重要的作用。此外,研究新的诊断应用,如DNA甲基化分析需要大量的培训和验证队列。目前,病理部门处理的大多数诊断病例都以福尔马林固定和石蜡包埋组织块(FFPE)的形式保存。因此,对从FFPE组织样品中提取高通量核酸的需求日益增长。虽然有专有的工具包,但它们价格昂贵,而且灵活性很小。在这里,我们提出了ht-HiTE,一种高通量实现最近发表的高效DNA提取协议。这种方法可以使用液体处理器手动和自动处理96孔板,提供两种纯化选择,并利用现成的试剂。最后,我们表明,与手工、基于试剂盒的方法相比,从ht-HiTE处理的DNA中获得的NGS和DNA甲基化微阵列数据具有相同的质量。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A cost-effective and scalable approach for DNA extraction from FFPE tissues.

Genomic profiling of cancer plays an increasingly vital role for diagnosis and therapy planning. In addition, research of novel diagnostic applications such as DNA methylation profiling requires large training and validation cohorts. Currently, most diagnostic cases processed in pathology departments are stored as formalin-fixed and paraffin embedded tissue blocks (FFPE). Consequently, there is a growing demand for high-throughput extraction of nucleic acids from FFPE tissue samples. While proprietary kits are available, they are expensive and offer little flexibility. Here, we present ht-HiTE, a high-throughput implementation of a recently published and highly efficient DNA extraction protocol. This approach enables manual and automated processing of 96-well plates with a liquid handler, offers two options for purification and utilizes off-the-shelf reagents. Finally, we show that NGS and DNA methylation microarray data obtained from DNA processed with ht-HiTE are of equivalent quality as compared to a manual, kit-based approach.

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来源期刊
Biology Methods and Protocols
Biology Methods and Protocols Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)
CiteScore
3.80
自引率
2.80%
发文量
28
审稿时长
19 weeks
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