Flow analysis and sorting of plant chromosomes

Vrána, J., Cápal, P., Šimková, H., Karafiátová, M., Čížková, J., Doležel, J.

Keywords: cell cycle synchronization, chromosome isolation, chromosome sorting, chromosome-specific DNA libraries, chromosome genomics, flow cytometry, high-molecular-weight DNA, genome sequencing, plant chromosomes, physical genome mapping
Abstract: Analysis and sorting of plant chromosomes (plant flow cytogenetics) is a special application of flow cytometry in plant genomics and its success depends critically on sample quality. This unit describes the methodology in a stepwise manner, starting with the induction of cell cycle synchrony and accumulation of dividing cells in mitotic metaphase, and continues with the preparation of suspensions of intact mitotic chromosomes, flow analysis and sorting of chromosomes, and finally processing of the sorted chromosomes. Each step of the protocol is described in detail as some procedures have not been used widely. Supporting histograms are presented as well as hints on dealing with plant material; the utility of sorted chromosomes for plant genomics is also discussed.
DOI: 10.1002/cpcy.9
Fulltext: contact IEB authors
IEB authors: Petr Cápal, Jana Čížková, Jaroslav Doležel, Miroslava Karafiá..., Hana Šimková, Jan Vrána