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Systematic hybrid LOH: a new method to reduce false positives and negatives during screening of yeast gene deletion libraries by David Alvaro; Ivana Sunjevaric; Robert J. D. Reid; Michael Lisby; David J. Stillman; Rodney Rothstein is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Abstract We have developed a new method, systematic hybrid loss of heterozygosity, to facilitate genomic screens utilizing the yeast gene deletion library. Screening is performed using hybrid diploid strains produced through mating the library haploids with strains from a different genetic background, to minimize the contribution of unpredicted recessive genetic factors present in the individual library strains. We utilize a set of strains where each contains a conditional centromere construct on one of the 16 yeast chromosomes that allows the destabilization and selectable loss of that chromosome. After mating a library gene deletion haploid to such a conditional centromere strain, which corresponds to the chromosome carrying the gene deletion, loss of heterozygosity (LOH) at the gene deletion locus can be generated in these otherwise hybrid diploids. The use of hybrid diploid strains permits complementation of any spurious recessive mutations in the library strain, facilitating attribution of the observed phenotype to the documented gene deletion and dramatically reducing false positive results commonly obtained in library screens. The systematic hybrid LOH method can be appli

Who reads Systematic hybrid LOH: a new method to reduce false positives and negatives during screening of yeast gene deletion libraries?

Author

David Alvaro; Ivana Sunjevaric; Robert J. D. Reid; Michael Lisby; David J. Stillman; Rodney Rothstein

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); John Wiley & Sons Inc.; [Chichester]; [New York]: John Wiley & Sons, ©1985-; Wiley; Hindawi Limited (ISSN 0749-503X)

Published

2006

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)