Details

Autor(en) / Beteiligte

• Dai, Zhizheng
• Ji, Hongyu
• Zheng, Anqi
• Huang, Ai-Long
• Tang, Kai-Fu

Titel

Protocol for isolating small cytosolic dsDNA from cultured murine cells

Ist Teil von

• STAR protocols, 2024-04, Vol.5 (2), p.102998-102998, Article 102998

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2024

Link zum Volltext

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• We recently identified a class of small cytosolic double-stranded DNA (scDNA) approximately 20–40 bp in size in human and mouse cells. Here, we present a protocol for scDNA isolation from cultured murine cells. We describe steps for cytosolic compartment separation, DNA isolation in the cytosolic fraction using phenol-chloroform extraction, and ethanol precipitation. We then detail procedures for denaturing purified cytosolic DNA through urea polyacrylamide gel electrophoresis and obtaining scDNA in the cytosolic DNA fraction via gel purification. For complete details on the use and execution of this protocol, please refer to Liu et al.1 [Display omitted] •Protocol for the isolation of scDNA from cultured murine cells•Steps to purify cytosolic DNA with phenol-chloroform extraction and ethanol precipitation•Steps to isolate scDNA from cytosolic DNA via urea polyacrylamide gel electrophoresis Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. We recently identified a class of small cytosolic double-stranded DNA (scDNA) approximately 20–40 bp in size in human and mouse cells. Here, we present a protocol for scDNA isolation from cultured murine cells. We describe steps for cytosolic compartment separation, DNA isolation in the cytosolic fraction using phenol-chloroform extraction, and ethanol precipitation. We then detail procedures for denaturing purified cytosolic DNA through urea polyacrylamide gel electrophoresis and obtaining scDNA in the cytosolic DNA fraction via gel purification.