Selecting Schizosaccharomyces pombe diploids

Karl Ekwall; Genevieve Thon

doi:10.1101/pdb.prot091702

Selecting Schizosaccharomyces pombe diploids

Karl Ekwall^*, Genevieve Thon

^*Corresponding author for this work

Functional Genomics

5 Citations (Scopus)

Abstract

Here we describe procedures for the selection of diploid Schizosaccharomyces pombe. ade6-M210/ade6-M216 heteroallelic complementation is widely used to select for Ade⁺ diploids. Such diploids will readily sporulate when starved of nitrogen. For some investigations, stable diploids are preferable (e.g., for genetic complementation tests), and in these cases mating an h⁻ strain with an h⁹⁰ mat2-Pi-102 strain can be used to prevent sporulation. When ade6-M210/ade6-M216 mutations impact on, or show synthetic interactions with, the gene of interest, two different auxotrophic markers can be used to select complementation.

Original language	English
Journal	Cold Spring Harbor Protocols
Volume	2017
Issue number	7
Pages (from-to)	537-540
Number of pages	4
ISSN	1940-3402
DOIs	https://doi.org/10.1101/pdb.prot091702
Publication status	Published - Jul 2017

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10.1101/pdb.prot091702

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abstract = "Here we describe procedures for the selection of diploid Schizosaccharomyces pombe. ade6-M210/ade6-M216 heteroallelic complementation is widely used to select for Ade+ diploids. Such diploids will readily sporulate when starved of nitrogen. For some investigations, stable diploids are preferable (e.g., for genetic complementation tests), and in these cases mating an h− strain with an h90 mat2-Pi-102 strain can be used to prevent sporulation. When ade6-M210/ade6-M216 mutations impact on, or show synthetic interactions with, the gene of interest, two different auxotrophic markers can be used to select complementation.",

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AB - Here we describe procedures for the selection of diploid Schizosaccharomyces pombe. ade6-M210/ade6-M216 heteroallelic complementation is widely used to select for Ade+ diploids. Such diploids will readily sporulate when starved of nitrogen. For some investigations, stable diploids are preferable (e.g., for genetic complementation tests), and in these cases mating an h− strain with an h90 mat2-Pi-102 strain can be used to prevent sporulation. When ade6-M210/ade6-M216 mutations impact on, or show synthetic interactions with, the gene of interest, two different auxotrophic markers can be used to select complementation.

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Selecting Schizosaccharomyces pombe diploids

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