Abstract
DNA cloned in yeast artificial chromosomes (YACs) is a valuable resource for functional experiments in cell culture as well as whole animal systems. Where the size or chimerism of a YAC clone are limiting factors it may be desirable to generate recombinant YAC clones. One such approach is based on mitotic recombination, and we describe the development of a methodology that allows multiple recombination cycles for serial reconstruction of overlapping YACs. This approach employs retrofitting with standard plasmid vectors, transfer of YACs to a common haploid host by Kar1 mating, and selection for recombination with 5-fluoro-orotic acid.
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© 2006 Humana Press Inc., Totowa NJ
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Markie, D., Jones, E., Ragoussis, J. (2006). Genomic Reconstruction by Serial Mitotic Recombination of Yeast Artificial Chromosomes. In: MacKenzie, A. (eds) YAC Protocols. Methods in Molecular Biology™, vol 349. Humana Press. https://doi.org/10.1385/1-59745-158-4:117
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DOI: https://doi.org/10.1385/1-59745-158-4:117
Publisher Name: Humana Press
Print ISBN: 978-1-58829-612-2
Online ISBN: 978-1-59745-158-1
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