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
{"title":"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。","authors":"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","doi":"10.1016/j.xpro.2024.103365","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103365"},"PeriodicalIF":1.3000,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11490689/pdf/","citationCount":"0","resultStr":"{\"title\":\"Protocol for the simultaneous isolation of DNA, RNA, and miRNA from a single archived Kaposi sarcoma biopsy.\",\"authors\":\"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\",\"doi\":\"10.1016/j.xpro.2024.103365\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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.</p>\",\"PeriodicalId\":34214,\"journal\":{\"name\":\"STAR Protocols\",\"volume\":\"5 4\",\"pages\":\"103365\"},\"PeriodicalIF\":1.3000,\"publicationDate\":\"2024-12-20\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11490689/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"STAR Protocols\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1016/j.xpro.2024.103365\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/10/4 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q4\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2024.103365","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/10/4 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
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.