Ekwall Karl, Thon Geneviève
Department of Biosciences and Nutrition, Karolinska Institute, Stockholm SE-141 83, Sweden;
Department of Biology, University of Copenhagen, Copenhagen DK-2200, Denmark.
Cold Spring Harb Protoc. 2017 Jul 5;2017(7):pdb.prot091694. doi: 10.1101/pdb.prot091694.
Here we provide methods for setting up standard crosses with strains. All strain genotypes and pedigrees should be recorded in a laboratory strain book. Matings between two haploid strains of interest are induced on solid medium poor in nitrogen. Usually, sporulation agar (SPA) plates are preferred, but for difficult matings it is advisable to try several mating media in parallel because one medium might allow for more efficient mating. Protoplast fusion can be used to produce zygotes from sterile mutants that fail to mate.
在这里,我们提供了用特定菌株进行标准杂交的方法。所有菌株的基因型和谱系都应记录在实验室菌株簿中。在氮含量低的固体培养基上诱导两个感兴趣的单倍体菌株之间进行交配。通常,孢子形成琼脂(SPA)平板是首选,但对于困难的交配,建议同时尝试几种交配培养基,因为一种培养基可能允许更有效的交配。原生质体融合可用于从不交配的无菌突变体中产生合子。