학술논문
Immunofluorescence microscopy-based detection of ssDNA foci by BrdU in mammalian cells
Document Type
Article
Source
STAR Protocols; December 2021, Vol. 2 Issue: 4
Subject
Language
ISSN
26661667
Abstract
DNA end resection converts broken ends of double-stranded DNA (dsDNA) to 3′-single-stranded DNA (3′-ssDNA). The extent of resection regulates DNA double-strand break (DSB) repair pathway choice and thereby genomic stability. Here, we characterize an optimized immunofluorescence (IF) microscopy-based protocol for measuring ssDNA in mammalian cells by labeling genomic DNA with 5-bromo-2′-deoxyuridine (BrdU). BrdU foci can be detected under non-denaturing conditions by anti-BrdU antibody, providing an accurate and reliable readout of DNA end resection in most mammalian cell lines.