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  • High-throughput genetic screening of meiotic commitment using fluorescence microscopy in Saccharomyces cerevisiae.

High-throughput genetic screening of meiotic commitment using fluorescence microscopy in Saccharomyces cerevisiae.

STAR protocols (2022-11-04)
Janardan N Gavade, Soni Lacefield
ABSTRACT

Simple genetic screens in budding yeast have identified many conserved meiotic regulators. However, the identification of genes involved in specific steps of meiosis may require a more complex genetic screen that allows visualization of meiosis. Here, we describe a high-throughput protocol using fluorescence microscopy to systematically screen an overexpression library to identify genes involved in meiotic commitment. We also explain how this protocol can be adapted for identifying proteins that function at different stages of meiosis. For complete details on the use and execution of this protocol, please refer to Gavade et al. (2022).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
myo-Inositol, ≥99%
Sigma-Aldrich
Glycine, suitable for electrophoresis, ≥99%
Sigma-Aldrich
L-Cysteine, 97%
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
L-Tyrosine, reagent grade, ≥98% (HPLC)