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

Authors

Larissa L. Scholte, Department of Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC 20052, USA. Electronic address: larissascholte@gwu.edu.
Justin Browne, Department of Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC 20052, USA.
David J. Nolan, Bioinfoexperts, LLC, Thibodaux, LA 70301, USA.
Peyton St John, Department of Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC 20052, USA.
Katherine Tracy, Department of Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC 20052, USA.
Rafaela S. Thur, Department of Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC 20052, USA.
Ghangzhao Li, Department of Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC 20052, USA.
Susanna L. Lamers, Bioinfoexperts, LLC, Thibodaux, LA 70301, USA.
Paige Bracci, Department of Epidemiology and Biostatistics, University of California, San Francisco, San Francisco, CA 94158, USA.
Michael S. McGrath, Department of Laboratory Medicine, Pathology, and Medicine, University of California, San Francisco, San Francisco, CA 94110, USA.
Jeffrey M. Bethony, Department of Microbiology, Immunology and Tropical Medicine, The George Washington University, Washington, DC 20052, USA. Electronic address: jbethony@gwu.edu.

Document Type

Journal Article

Publication Date

10-4-2024

Journal

STAR protocols

Volume

5

Issue

4

DOI

10.1016/j.xpro.2024.103365

Keywords

Cancer; Gene Expression; Molecular Biology; Sequencing

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.

APA Citation

Scholte, Larissa L.; Browne, Justin; Nolan, David J.; St John, Peyton; Tracy, Katherine; Thur, Rafaela S.; Li, Ghangzhao; Lamers, Susanna L.; Bracci, Paige; McGrath, Michael S.; and Bethony, Jeffrey M., "Protocol for the simultaneous isolation of DNA, RNA, and miRNA from a single archived Kaposi sarcoma biopsy" (2024). GW Authored Works. Paper 5875.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/5875

Department

Microbiology, Immunology, and Tropical Medicine