ABSTRACT
Methods of chromosome analysis and sorting by flow cytometry are increasingly used in plant genomics. Protocols for flow cytometry are provided by J. Dolezel which was published by Dolezel et al. and Vrana et al. Flow cytometry can be used for rapid classification of isolated chromosomes according to their relative DNA content. A general outline of the procedure for flow cytometric analysis and sorting of plant chromosomes consists of the following steps: induction of cell cycle synchrony and accumulation of dividing cells in metaphase, preparation of suspensions of intact chromosomes, flow analysis and sorting, and processing of flow-sorted chromosomes. During the initial flow karyotyping experiment in a given plant species or line, it is useful to sort chromosomes onto a microscopic slide to determine the chromosome content of individual populations on a flow karyotype, and to examine the purity that may be achieved when sorting specific chromosomes.
