High-throughput transformation of Saccharomyces cerevisiae using liquid handling robots
AffiliationUniv Arizona, Dept Mol & Cellular Biol
Univ Arizona, Funct Genom Core Facil
MetadataShow full item record
PublisherPUBLIC LIBRARY SCIENCE
CitationHigh-throughput transformation of Saccharomyces cerevisiae using liquid handling robots 2017, 12 (3):e0174128 PLOS ONE
Rights© 2017 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License.
Collection InformationThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at email@example.com.
AbstractSaccharomyces cerevisiae (budding yeast) is a powerful eukaryotic model organism ideally suited to high-throughput genetic analyses, which time and again has yielded insights that further our understanding of cell biology processes conserved in humans. Lithium Acetate (LiAc) transformation of yeast with DNA for the purposes of exogenous protein expression (e.g., plasmids) or genome mutation (e.g., gene mutation, deletion, epitope tagging) is a useful and long established method. However, a reliable and optimized high throughput transformation protocol that runs almost no risk of human error has not been described in the literature. Here, we describe such a method that is broadly transferable to most liquid handling high-throughput robotic platforms, which are now commonplace in academic and industry settings. Using our optimized method, we are able to comfortably transform approximately 1200 individual strains per day, allowing complete transformation of typical genomic yeast libraries within 6 days. In addition, use of our protocol for gene knockout purposes also provides a potentially quicker, easier and more cost-effective approach to generating collections of double mutants than the popular and elegant synthetic genetic array methodology. In summary, our methodology will be of significant use to anyone interested in high throughput molecular and/or genetic analysis of yeast.
NoteOpen Access Journal.
VersionFinal published version
SponsorsUniversity of Arizona; NIH [R01 GM1145664-01A1]