从单个存档卡波西肉瘤活检组织中同时分离 DNA、RNA 和 miRNA 的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-10-04 DOI:10.1016/j.xpro.2024.103365

Larissa L S Scholte, Justin Browne, David J Nolan, Peyton St John, Katherine Tracy, Rafaela S Thur, Ghangzhao Li, Susanna L Lamers, Paige Bracci, Michael S McGrath, Jeffrey M Bethony

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引用次数: 0

摘要

卡波西肉瘤（KS）打孔活检为提取核酸带来了独特的挑战，而核酸在 RNAlater 中的长期稳定又加剧了这种挑战。在此，我们介绍一种同时从单个 KS 穿孔活检组织中分离 DNA、RNA 和 miRNA 的方案。我们详细介绍了准备试剂和用品、使用手工和机械方法破坏 KS 组织、分离 DNA 和总 RNA、评估核酸质量以及长期储存核酸的步骤。

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Protocol for the simultaneous isolation of DNA, RNA, and miRNA from a single archived Kaposi sarcoma biopsy.

Kaposi sarcoma (KS) punch biopsies present unique challenges for extracting nucleic acids, which can be exacerbated by their long-term stabilization in RNAlater. Here, we present a protocol for simultaneously isolating DNA, RNA, and miRNA from a single KS punch biopsy. We detail the steps for preparing reagents and supplies, disrupting KS tissue using manual and mechanical methods, isolating DNA and total RNA, evaluating nucleic acid quality, and storing nucleic acids long-term.

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