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

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

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.

Keywords: Cancer; Gene Expression; Molecular Biology; Sequencing.

MeSH terms

  • Biopsy / methods
  • DNA / genetics
  • DNA / isolation & purification
  • Humans
  • MicroRNAs* / analysis
  • MicroRNAs* / genetics
  • MicroRNAs* / isolation & purification
  • RNA / analysis
  • RNA / genetics
  • RNA / isolation & purification
  • Sarcoma, Kaposi* / diagnosis
  • Sarcoma, Kaposi* / genetics
  • Sarcoma, Kaposi* / pathology

Substances

  • MicroRNAs
  • DNA
  • RNA