Šafář, J., Šimková, H., Kubaláková, M., Číhalíková, J., Suchánková, P., Bartoš, J., Doležel, J.
CYTOGENETIC AND GENOME RESEARCH 129: 211-223, 2010

Keywords: BAC library, Chromosome, DNA markers, Flow sorting, Physical mapping, Wheat
Abstract: The large bread wheat genome (1C - 17 Gbp) contains a preponderance of repetitive DNA and the species is polyploid. These characteristics together serve to hamper the molecular analysis of the wheat genome. Its complexity can, however, be reduced by using flow cytometry to isolate individual chromosomes, and these can be exploited to construct chromosome-specific BAC libraries. Such libraries simplify the task of physical map construction, positional cloning and the targeted development of genetic markers. Rapid improvements in the efficiency and cost of DNA sequencing provide an opportunity to contemplate sequencing the wheat genome by preparing sequence-ready physical maps for each chromosome or chromosome arm in turn. The quality of the chromosome-specific libraries depends on their chromosome coverage and the mean insert size. First-generation libraries suffered from a relatively low mean insert size, but improvements to the protocol have generated a second wave of libraries with a significantly increased mean insert size and better chromosome coverage. Each chromosome (arm)-specific library is composed of a manageable number of clones, and so represents a practical tool in the area of wheat genomics.
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IEB authors: Jan Bartoš, Jaroslav Doležel, Jan Šafář, Hana Šimková, Pavla Suchánková