RT Journal Article
SR Electronic
T1 Automation of yeast spot assays using an affordable liquid handling robot
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.07.16.500326
DO 10.1101/2022.07.16.500326
A1 Taguchi, Shodai
A1 Suda, Yasuyuki
A1 Irie, Kenji
A1 Ozaki, Haruka
YR 2022
UL http://biorxiv.org/content/early/2022/07/18/2022.07.16.500326.abstract
AB The spot assay of the budding yeast Saccharomyces cerevisiae is an experimental method that is used to evaluate the effect of genotypes, medium conditions, and environmental stresses on cell growth and survival. However, there has yet to be an affordable solution for the automation of spot assays. To make reproducible spot assay experiments widely available, we have automated the plate-based yeast spot assay of budding yeast using Opentrons OT-2 (OT-2), an affordable liquid-handling robot, and a flatbed scanner. We prepared a 3D-printed mount for the Petri dish to allow for precise placement of the Petri dish inside the OT-2. To account for the uneven height of the agar plates, which were made by human hands, we devised a method to adjust the z-position of the pipette tips which is based on the weight of each agar plate. During the incubation of the agar plates, a flatbed scanner was used to automatically take images of the agar plates over time, allowing researchers to quantify and compare the cell density within the spots at optimal time points a posteriori. Furthermore, the accuracy of the newly developed automated spot assay was verified by performing spot assays with human experimenters and the OT-2 and quantifying the yeast-grown area of the spots. This study will contribute to the introduction of automated spot assays and the automated acquisition of growth processes in conventional laboratories that are not adapted for high-throughput laboratory automation.Competing Interest StatementThe authors have declared no competing interest.