Vrána, J., Cápal, P., Číhalíková, J., Kubaláková, M., Doležel, J.
Plant Cytogenetics: Methods and Protocols : 119-134, 2016

Keywords: Cell cycle synchronization , Chromosome isolation , Cytogenetic stocks , FISH , FISHIS , Flow cytometry and sorting , Metaphase accumulation , Plants
Abstract: Nuclear genomes of many important plant species are tremendously complicated to map and sequence. The ability to isolate single chromosomes, which represent small units of nuclear genome, is priceless in many areas of plant research including cytogenetics, genomics, and proteomics. Flow cytometry is the only technique which can provide large quantities of pure chromosome fractions suitable for downstream applications including physical mapping, preparation of chromosome-specifi c BAC libraries, sequencing, and optical mapping. Here, we describe step-by-step procedure of preparation of liquid suspensions of intact mitotic metaphase chromosomes and their fl ow cytometric analysis and sorting.
DOI: 10.1007/978-1-4939-3622-9_10
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IEB authors: Petr Cápal, Jaroslav Doležel, Jan Vrána