Protocol to investigate cell cycle-dependent modes of micronucleus formation using dual-pulse nascent DNA labeling

Here, we present a protocol to study the contribution of individual cell-cycle stages to genome instability mechanisms using dual-pulse nascent DNA labeling. We describe steps for sample treatment, immunofluorescence staining, imaging, and image processing. This protocol enables classification of cells according to their cell-cycle phase at the time of drug treatment of interest. It is valuable for investigating cell cycle-dependent treatment effects and is compatible with immunofluorescence-based analyses. For complete details on the use and execution of this protocol, please refer to Pepe et al.¹ and Duardo et al.².