title: Improving detection of rare biological events in high-throughput screens.
abstract: The success of high-throughput screening (HTS) strategies depends on the effectiveness of both normalization methods and study design. We report comparisons among normalization methods in two titration series experiments. We also extend the results in a third experiment with two differently designed but otherwise identical screens: compounds in replicate plates were either placed in the same well locations or were randomly assigned to different locations. Best results were obtained when randomization was combined with normalization methods that corrected for within-plate spatial bias. We conclude that potent, reliable, and accurate HTS requires replication, randomization design strategies, and more extensive normalization than is typically done and that formal statistical testing is desirable. The Statistics and dIagnostic Graphs for HTS (SIGHTS) Microsoft Excel Add-In software is available to conduct most analyses reported here.
authors.affiliation: McGill University and Genome Quebec Innovation Centre, Montreal, Quebec, Canada Department of Human Genetics, McGill University, Montreal, Quebec, Canada.
authors.forename: Carl
authors.initials: C
authors.lastname: Murie
authors.affiliation: Equipe Criblage pour des Molécules Bio-Actives (CMBA), U1038 INSERM/CEA/UJF, CEA Grenoble, Grenoble Cedex 09, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Barette
authors.affiliation: McGill University and Genome Quebec Innovation Centre, Montreal, Quebec, Canada.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Button
authors.affiliation: Equipe Criblage pour des Molécules Bio-Actives (CMBA), U1038 INSERM/CEA/UJF, CEA Grenoble, Grenoble Cedex 09, France Institut Albert Bonniot, CRI INSERM/UJF U823, Team 3 "Polarity, Development and Cancer," Rond-point de la Chantourne, La Tronche Cedex, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Lafanechère
authors.affiliation: McGill University and Genome Quebec Innovation Centre, Montreal, Quebec, Canada Department of Human Genetics, McGill University, Montreal, Quebec, Canada robert.nadon@mcgill.ca.
authors.forename: Robert
authors.initials: R
authors.lastname: Nadon
mesh_terms: D001331:Automation; D055808:Drug Discovery / Q000379:methods* / Q000592:standards*; D006367:HeLa Cells; D057166:High-Throughput Screening Assays / Q000592:standards*; D006801:Humans; D015233:Models, Statistical; D011786:Quality Control; D015203:Reproducibility of Results; D054852:Small Molecule Libraries
chemical_list: D054852:Small Molecule Libraries
keywords: normalization; randomization; statistical analysis; study design