A method for quantifying sporulation efficiency and isolating meiotic progeny in non-GMO strains of Saccharomyces cerevisiae.
Amy SirrMartin S TimourGareth A CromieMichelle TangAimeé M DudleyPublished in: Yeast (Chichester, England) (2022)
Meiotic mapping, a linkage-based method for analyzing the recombinant progeny of a cross, has long been a cornerstone of genetic research. The yeast Saccharomyces cerevisiae is a powerful system because it is possible to isolate and cultivate the four products (spores) of a single meiotic event. However, the throughput of this process has historically been limited by the process of identifying tetrads in a heterogeneous population of vegetative cells, tetrads, and dyads followed by manual separation (dissection) of the spores contained in a tetrad. To date, methods that facilitate high throughput characterization and isolation of meiotic progeny have relied on genetic engineering. Here, we characterize the ability of the fluorescent dye DiBAC 4 (5) to stain yeast tetrads and dyads as well as to adhere to spores following bulk tetrad disruption. Applications include quantitative assays of sporulation rates and efficiency by flow cytometry as well as enrichment of intact tetrads, dyads, or disrupted spores by fluorescence-activated cell sorting in strains that have not been genetically modified.
Keyphrases
- saccharomyces cerevisiae
- high throughput
- flow cytometry
- genome wide
- single cell
- escherichia coli
- induced apoptosis
- high resolution
- cell cycle arrest
- copy number
- bacillus subtilis
- quantum dots
- cell therapy
- dna methylation
- cell death
- liquid chromatography
- cell proliferation
- mass spectrometry
- pi k akt
- hepatitis c virus
- hiv testing
- aqueous solution