Yeast Resources at the WIMM

The WIMM houses several core pieces of equipment and reagents for both classical genetic analysis and high-throughput/systems-level analysis in budding yeast (Saccharomyces cerevisiae). For classical and low-throughput genetics, molecular and cell biology, expertise and a large range of cloning, tagging and expression vectors are available. Specific additional technology and resources available locally include:

Yeast tetrad dissection microscope

Following mating of haploid yeast strains and subsequent sporulation, this microscope allows dissection of yeast tetrads, asci containing four spores that are meiotic progeny. This is essential for classical, low-throughput genetic analysis, synthetic lethality analysis and a ‘clean’ way of creating double/multiple mutant strains through genetic crosses.

Singer ROTA HDA Robot

yeast
This robot (www.singerinstruments.com) can is able to replicate high-density arrays of yeast strains, of up to 1536 colonies per plate starting with 96-well microtitre plates or low-density arrays of strains plated on agar. The speed of the robot coupled with the high density of the arrays enables replica plating of the entire yeast gene-deletion strain collection (approx. 4800 strains, see below), in only a few minutes. In addition, Synthetic Genetic Analysis (SGA) and other systems-biology and genome-wide genetic techniques (synthetic dosage lethality and suppression analysis) are made possible by this high-density, high-throughput technology. These approaches enable the creation of genetic interaction networks, large-scale 2-hybrid analysis as well as phenotypic and chemical-genetic analysis.

Yeast deletion library

This collection was developed by the Saccharomyces Genome Deletion Project, and includes knock-out strains covering 96% of the yeast non-essential genome (approx. 4800 strains). The distinct tags flanking the disruption cassette, also known as "molecular barcodes", uniquely identify each strain allowing phenotypic analysis approaches to be performed on genome-wide level (synthetic lethality analysis by microarray, or SLAM). At the WIMM we have the haploid deletion collection in an ‘a-mating type’ background.

Yeast ORF library

The Yeast ORF library consists of over 4,900 yeast open reading frames (ORFs) that have been adapted for use with the Gateway system. The ORFs are expressed as a C-terminal fusion protein (6xHis-HA, cleavable) under control of a regulatable GAL promoter.

Additional reading:

Ooi SL et al. Global synthetic-lethality analysis and yeast functional profiling. Trends Genet. 2006 22:56-63.