The analysis of the complex genome of common wheat (Triticum aestivum, 2n = 6x = 42, genome formula AABBDD) is hampered by its large size ( approximately 17 000 Mbp) and allohexaploid nature. In order to simplify its analysis, we developed a generic strategy for dissecting such large and complex genomes into individual chromosomes. Chromosome 3B was successfully sorted by flow cytometry and cloned into a bacterial artificial chromosome (BAC), using only 1.8 million chromosomes and an adapted protocol developed for this purpose. The BAC library (designated as TA-3B) consists of 67 968 clones with an average insert size of 103 kb. It represents 6.2 equivalents of chromosome 3B with 100% coverage and 90% specificity as confirmed by genetic markers. This method was validated using other chromosomes and its broad application and usefulness in facilitating wheat genome analysis were demonstrated by target characterization of the chromosome 3B structure through cytogenetic mapping. This report on the successful cloning of flow-sorted chromosomes into BACs marks the integration of flow cytogenetics and genomics and represents a great leap forward in genetics and genomic analysis.

Laboratory of Molecular Cytogenetics and Cytometry Institute of Experimental Botany Sokolovska 6 CZ 77200 Olomouc Czech Republic

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Flow Cytometric Analysis and Sorting of Plant Chromosomes

Flow karyotyping of wheat-Aegilops additions facilitate dissecting the genomes of Ae. biuncialis and Ae. geniculata into individual chromosomes

Flow sorting of C-genome chromosomes from wild relatives of wheat Aegilops markgrafii, Ae. triuncialis and Ae. cylindrica, and their molecular organization

High-throughput physical map anchoring via BAC-pool sequencing

Genome-wide analysis of repeat diversity across the family Musaceae

Chromosomes in the flow to simplify genome analysis

BAC libraries from wheat chromosome 7D: efficient tool for positional cloning of aphid resistance genes

Integration of genetic and physical maps of the chickpea (Cicer arietinum L.) genome using flow-sorted chromosomes

A first survey of the rye (Secale cereale) genome composition through BAC end sequencing of the short arm of chromosome 1R

Coupling amplified DNA from flow-sorted chromosomes to high-density SNP mapping in barley

A novel resource for genomics of Triticeae: BAC library specific for the short arm of rye (Secale cereale L.) chromosome 1R (1RS)

Chromosome-based genomics in the cereals

Dissection of the nuclear genome of barley by chromosome flow sorting

Chromosome sorting in tetraploid wheat and its potential for genome analysis

Construction of a subgenomic BAC library specific for chromosomes 1D, 4D and 6D of hexaploid wheat