Šafář, J., Šimková, H., Doležel, J.
Plant Cytogenetics: Methods and Protocols : 135-149, 2016

Keywords: BAC library , BAC vector , Chromosomes , DNA cloning , High molecular weight (HMW) DNA , Pulse-fi eld gel electrophoresis (PFGE) , Clone
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 purifi ed by fl ow cytometric sorting. Chromosome-specifi c BAC libraries facilitate positional gene cloning, physical mapping, and sequencing in complex plant genomes.
DOI: 10.1007/978-1-4939-3622-9_11
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IEB authors: Jaroslav Doležel, Jan Šafář, Hana Šimková