title: Flow Analysis and Sorting of Plant Chromosomes.
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. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Institute of Experimental Botany, Center of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Vrána
authors.affiliation: Institute of Experimental Botany, Center of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Cápal
authors.affiliation: Institute of Experimental Botany, Center of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic.
authors.forename: Hana
authors.initials: H
authors.lastname: Šimková
authors.affiliation: Institute of Experimental Botany, Center of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic.
authors.forename: Miroslava
authors.initials: M
authors.lastname: Karafiátová
authors.affiliation: Institute of Experimental Botany, Center of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic.
authors.forename: Jana
authors.initials: J
authors.lastname: Čížková
authors.affiliation: Institute of Experimental Botany, Center of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic.
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Doležel
mesh_terms: D032461:Chromosomes, Plant / Q000378:metabolism*; D018744:DNA, Plant / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D018519:Meristem / Q000166:cytology / Q000187:drug effects; D008677:Metaphase / Q000187:drug effects; D008970:Molecular Weight; D009609:Nitrous Oxide / Q000494:pharmacology; D010944:Plants / Q000235:genetics*; D040901:Proteomics; D012639:Seeds / Q000187:drug effects
chemical_list: D018744:DNA, Plant; D009609:Nitrous Oxide
keywords: cell cycle synchronization; chromosome genomics; chromosome isolation; chromosome sorting; chromosome-specific DNA libraries; flow cytometry; genome sequencing; high-molecular-weight DNA; physical genome mapping; plant chromosomes