title: Construction of BAC Libraries from Flow-Sorted Chromosomes.
abstract: Cloned DNA libraries in bacterial artificial chromosome (BAC) are the most widely used form of large-insert DNA libraries. BAC libraries are typically represented by ordered clones derived from genomic DNA of a particular organism. In the case of large eukaryotic genomes, whole-genome libraries consist of a hundred thousand to a million clones, which make their handling and screening a daunting task. The labor and cost of working with whole-genome libraries can be greatly reduced by constructing a library derived from a smaller part of the genome. Here we describe construction of BAC libraries from mitotic chromosomes purified by flow cytometric sorting. Chromosome-specific BAC libraries facilitate positional gene cloning, physical mapping, and sequencing in complex plant genomes.
authors: Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany, Šlechtitelů 31, 783 71, Olomouc, Czech Republic. safar@ueb.cas.cz.
authors: Jan
authors: J
authors: Šafář
authors: Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany, Šlechtitelů 31, 783 71, Olomouc, Czech Republic.
authors: Hana
authors: H
authors: Šimková
authors: Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany, Šlechtitelů 31, 783 71, Olomouc, Czech Republic.
authors: Jaroslav
authors: J
authors: Doležel
mesh_terms: D022202:Chromosomes, Artificial, Bacterial; D032461:Chromosomes, Plant; D005434:Flow Cytometry / Q000379:methods*; D018745:Genome, Plant; D015698:Genomic Library; D010944:Plants / Q000235:genetics*
keywords: BAC library; BAC vector; Chromosomes; Clone; DNA cloning; High molecular weight (HMW) DNA; Pulse-field gel electrophoresis (PFGE)