STAR Protocols· 2026Q2
Protocol for quantifying chromatin DNA replication by multiplex labeling and single-molecule localization microscopy
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- Q2SCImago
- 2026year
Short summary
A new protocol enables multiplex labeling and single-molecule localization microscopy (SMLM) to quantify DNA replication signals in synchronized mammalian cells.
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Key points
- Provides a step-by-step protocol for quantifying chromatin DNA replication.
- Utilizes cell-cycle synchronization and nucleoside analog labeling.
- Employs dual-color STORM imaging for high-resolution detection.
- Applies SR-Tesseler for cluster analysis of replication signals.
AI-generated from the title and abstract; the full text is not read.
Abstract
We present a protocol for imaging and quantifying chromatin DNA replication-associated signals in synchronized mammalian cells. We describe steps for cell-cycle enrichment, nucleoside analog-based metabolic labeling, and fluorescence detection. We then detail the procedure for dual-color STORM imaging and SR-Tesseler-based cluster analysis. It can be adapted to distinct questions in DNA replication biology by modifying synchronization schemes, pulse durations, labeling strategies, fluorescence detection channels, or analysis parameters. For complete details on the use and execution of this protocol, please refer to Zhang et al. 1
The authors' abstract, as published at the source. STAR Protocols, 2026 · DOI ↗
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BiophysicsBiochemistry, Genetics and Molecular Biology