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High-throughput phenotypic testing is increasingly important for exploring the biology of bacteria, fungi, yeasts, and animal cell lines such as human cancer cells.Just as DNA microarrays and proteomic technologies have made it possible to assay the expression level of thousands of genes or proteins all a once, phenotype microarrays (PMs) make it possible to quantitatively measure thousands of ...
High-throughput screening (HTS) is a method for scientific discovery especially used in drug discovery and relevant to the fields of biology, materials science [1] and chemistry. [ 2 ] [ 3 ] Using robotics , data processing/control software, liquid handling devices, and sensitive detectors, high-throughput screening allows a researcher to ...
Nevertheless, the implementation of Sanger sequencing for decoding DNA-encoded chemical libraries in high-throughput fashion was the first to be described. [18] After selection and PCR amplification of the DNA-tags of the library compounds, concatamers containing multiple coding sequences were generated and ligated into a vector .
High-content screening where changes in the expression of several proteins can be simultaneously monitored is also often used. [9] [10] High-content imaging of dye-labeled cellular components can also reveal effects of compounds on cell cultures in vitro, distinguishing the phenotypic effects of a broad variety of drugs. [11]
Early studies in Caenorhabditis elegans [1] and Drosophila melanogaster [2] [3] saw large-scale, systematic loss of function (LOF) screens performed through saturation mutagenesis, demonstrating the potential of this approach to characterise genetic pathways and identify genes with unique and essential functions.
The choice of a chemical library is less stringent than other high-throughput screening strategies owing to the lack of functional readouts, which would otherwise require deconvolution of the source compound that generates biological activity. Thus, the typical range for AS-MS is 400-3,000 compounds per pool. [20]
Unlike high-content analysis, high-content screening implies a level of throughput which is why the term "screening" differentiates HCS from HCA, which may be high in content but low in throughput. In high content screening, cells are first incubated with the substance and after a period of time, structures and molecular components of the cells ...
Similar to classical genetic screens in the past, large-scale RNAi surveys success depends on a careful development of phenotypic assays and their interpretation. [9] In Drosophila , RNAi has been applied in cultured cells or in vivo to investigate gene functions and to effect the function of single genes on a genome-wide scale.