title: Novel method for the high-throughput processing of slides for the comet assay.
abstract: Single cell gel electrophoresis (the comet assay), continues to gain popularity as a means of assessing DNA damage. However, the assay's low sample throughput and laborious sample workup procedure are limiting factors to its application. "Scoring", or individually determining DNA damage levels in 50 cells per treatment, is time-consuming, but with the advent of high-throughput scoring, the limitation is now the ability to process significant numbers of comet slides. We have developed a novel method by which multiple slides may be manipulated, and undergo electrophoresis, in batches of 25 rather than individually and, importantly, retains the use of standard microscope comet slides, which are the assay convention. This decreases assay time by 60%, and benefits from an electrophoresis tank with a substantially smaller footprint, and more uniform orientation of gels during electrophoresis. Our high-throughput variant of the comet assay greatly increases the number of samples analysed, decreases assay time, number of individual slide manipulations, reagent requirements and risk of damage to slides. The compact nature of the electrophoresis tank is of particular benefit to laboratories where bench space is at a premium. This novel approach is a significant advance on the current comet assay procedure.
authors.affiliation: Oxidative Stress Group, University of Leicester, Leicester, LE2 7LX, UK.
authors.forename: Mahsa
authors.initials: M
authors.lastname: Karbaschi
authors.affiliation: Oxidative Stress Group, University of Leicester, Leicester, LE2 7LX, UK.
authors.forename: Marcus S
authors.initials: MS
authors.lastname: Cooke
mesh_terms: D020552:Comet Assay / Q000379:methods*; D004247:DNA / Q000235:genetics* / Q000648:ultrastructure*; D004249:DNA Damage / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing; D012371:Robotics / Q000379:methods*
chemical_list: D004247:DNA