从存档的卡波西肉瘤活检样本中同时分离DNA、RNA和miRNA的方案。

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

作者信息

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

机构信息

Department of Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC 20052, USA.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103365. doi: 10.1016/j.xpro.2024.103365. Epub 2024 Oct 4.

DOI:10.1016/j.xpro.2024.103365

PMID:39368095

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11490689/

Abstract

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.

摘要

卡波西肉瘤（KS）穿刺活检在提取核酸方面存在独特挑战，而其在RNA later中进行长期保存会使这些挑战进一步加剧。在此，我们展示了一种从单个KS穿刺活检样本中同时分离DNA、RNA和miRNA的方案。我们详细介绍了制备试剂和耗材、使用手动和机械方法破坏KS组织、分离DNA和总RNA、评估核酸质量以及长期保存核酸的步骤。