Chromosome conformation capture of mitotic chromosomes

Cápal P.
Plant cytogenetics and cytogenomics: Methods and Protocols 2672: 485-500, 2023

Keywords: Chromatin interaction, 3D architecture, Hi-C, Flow sorting, Plant chromosomes
Abstract: Despite more than a century of intensive study of mitotic chromosomes, their three-dimensional organiza-tion remains enigmatic. The last decade established Hi-C as a method of choice for study of spatial genome-wide interactions. Although its utilization has been focused mainly on studying genomic interactions in interphase nuclei, the method can be also successfully applied to study 3D architecture and genome folding in mitotic chromosomes. However, obtaining sufficient number of mitotic chromosomes as an input material and effective coupling with Hi-C method is challenging in plant species. An elegant way to overcome hindrances with obtaining a pure mitotic chromosome fraction is their isolation via flow cyto-metric sorting. This chapter presents a protocol describing plant sample preparation for chromosome conformation studies, for flow-sorting of plant mitotic metaphase chromosomes and for the Hi-C procedure.
DOI: 10.1007/978-1-0716-3226-0_29
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IEB authors: Petr Cápal