Chromosome flow sorting and physical mapping

Doležel, Jaroslav; Kubaláková, Marie; Bartoš, Jan; Macas, Jiří
In The Handbook of Plant Genome Mapping. Genetic and Physical Mapping. Weinheim : Wiley-VCH Verlag GmbH & Co. KGaA : 151-171, 2005

Klíčová slova: Flow cytogenetics; genome mapping
Abstrakt: Flow cytometry is a powerful tool for quantitative analysis and purification of mitotic chro-mosomes. The analysis is performed at high rates, typically 102 – 103 / sec, and large quanti-ties of chromosomes stained by a DNA-binding fluorochrome are classified according to fluo-rescence intensity which reflects their relative DNA content. The resulting distributions of DNA content are called flow karyotypes. Ideally, each chromosome on a flow karyotype is represented by a single well discriminated peak. Any chromosome or a group of chromo-somes that may be discriminated may also be purified by sorting. Two different chromosomes may be sorted simultaneously. Flow karyotyping has been found suitable for quantitative de-tection of specific structural and numerical chromosome changes. Large numbers of chromo-somes that may be sorted onto microscopic slide facilitate high resolution physical mapping using FISH or PRINS and discovery of rare structural changes. PCR with chromosomes sorted into reaction tubes allows physical mapping of short and single copy targets; the use of deletion and translocation chromosomes enables subchromosomal localization of mapped sequences. DNA of sorted chromosomes is suitable for cloning and preparation of chromo-some-specific DNA libraries. Short insert libraries have been used for targeted isolation of molecular markers from specific genome regions. It is expected that the availability of chro-mosome- and chromosome-arm specific BAC libraries harbouring large inserts will greatly simplify development of physical contig maps and gene isolation in plant species possessing large and complex genomes.
Autoři z ÚEB: Jan Bartoš, Jaroslav Doležel