DNA fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells

STAR Protocols

Judit Jimenez Sainz¹  Gemma Moore²  Ryan B. Jensen² 

[1] Corresponding author;Department of Therapeutic Radiology, Yale University School of Medicine, 15 York Street, New Haven, CT 06520, USA;

关键词: Cell Biology;    Molecular Biology;    Single-molecule Assays;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: DNA fiber combing is a versatile technique that provides insight into replication fork dynamics at single-molecule resolution. DNA fibers are bound to silanized coverslips and combed, which straightens and aligns the fibers along a single axis. Here, we present a DNA fiber combing protocol that does not use commercial kits; we detail the steps to prepare all materials, reagents, and silanized coverslips. We describe the use of DLD-1 cells, but the protocol is amenable to other cell types.

【 授权许可】

Unknown