The ROTOR is the fastest and most powerful colony manipulation robot in the world. It is essential for any large-scale microbial functional or chemical genomic studies.
The Stinger is an automated modular extension to the ROTOR HDA robot for easy, ultra-fast rearrangement of high-density library arrays of yeast, other fungi, bacteria, and algae from colonies grown on solid agar or in liquid culture. It is designed for the picking and spotting of user-specified colonies from high-density arrays onto solid agars in 96-, 384-, 1536-, and 6144-density format.
Synthetic Genetic Array (SGA) analysis involves the manipulation of high-density yeast arrays to identify synthetic genetic interactions. In a typical SGA screen, a query strain is systematically crossed to an ordered array of deletion mutants, such that the meiotic progeny harboring both gene mutations can be assayed for growth defects. Identifying these interactions allows for the construction of large genetic networks, which reveal functional dependency and pathway redundancy in yeast.
The yeast 2-hybrid (Y2H) assay is a well-established technique to detect protein-protein interactions. This is an extremely powerful tool for researchers and is often used alongside one or two other methods to examine the multitude of interactions that take place in cells. The assay is straightforward to perform and generates high quality results in a short amount of time.
We are carrying out broad genetic screens in the green alga Chlamydomonas reinhardtii, for which the ROTOR HDA technology would be very useful. However, Chlamydomonas differs from budding yeast in cell size, growth rate, colony morphology and adhesiveness, and light responsiveness. Therefore, it was important to test specifically if the ROTOR HDA technology would work with Chlamydomonas.
Chemical genetics is the study of genes through small-molecule perturbation. Chemical genetic screening is a phenotypic screening methodology that systematically tests the efficacy of thousands of small molecules simultaneously. It is superior to target-based screening methodology for drug discovery because chemical genetic screening is unbiased and screens chemicals directly in complex cellular environment. Therefore, chemical genetic screening is an efficient way to discover and validate new drugable targets and identify potentially efficacious therapeutics.
E. coli Synthetic Genetic Array (eSGA) analysis involves the manipulation of high-density bacteria arrays to monitor bacterial genetic interactions. In a typical eSGA screen, a query strain is systematically conjugated to a genome-wide arrayed collection of single gene mutants. After rounds of robotic pinning and duel marker selection, E. coli strains harboring both gene mutations can be assayed for growth defects. Systematic and quantitative identification of genetic interactions allows the construction of large genetic networks, revealing functional dependency and pathway redundancy in E. coli.
We evaluate here the use of The Stinger single colony picker for agar to agar transfer of individual colonies of Chlamydomonas from 1536 and 6144 array densities. Specifically, we assess the accuracy with which The Stinger selects the desired colony within the source array, the compatibility of the pin material and transfer speed with the viability of Chlamydomonas cells, and the extent of cross-contamination between cells in the desired colony and those of the surrounding colonies.