Two-parameter single molecule analysis for measurement of chromatin mobility

This protocol provides a two-parameter analysis of live-cell Single Molecule Tracking (SMT) trajectories of Halo-tagged histone and unveils a chromatin mobility landscape composed of five chromatin types ranging from low to high mobility. When the analysis is applied to Halo-tagged, chromatin-binding proteins, it associates chromatin interaction properties with known functions in a way that previously used SMT parameters did not. For complete details on the use and execution of this protocol, please refer to Lerner et al., 2020.

本方案针对经Halo标签标记的组蛋白的活细胞单分子追踪（Single Molecule Tracking, SMT）轨迹提供了双参数分析方法，并揭示了由五种迁移速率从低到高的染色质类型构成的染色质运动图谱。当该分析方法应用于经Halo标签标记的染色质结合蛋白时，可将染色质相互作用特性与已知功能关联起来，这是过往所使用的SMT参数无法达成的。如需了解该方案的完整使用与实施细节，请参阅Lerner等人2020年的研究成果。