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• W2294418165 abstract "Sort-seq is an effective approach for simultaneous activity measurements in a large-scale library, combining flow cytometry, deep sequencing, and statistical inference. Such assays enable the characterization of functional landscapes at unprecedented scale for a wide-reaching array of biological molecules and functionalities in vivo. Applications of sort-seq range from footprinting to establishing quantitative models of biological systems and rational design of synthetic genetic elements. Nearly as diverse are implementations of this technique, reflecting key design choices with extensive impact on the scope and accuracy the results. Yet how to make these choices remains unclear. Here we investigate the effects of alternative sort-seq designs and inference methods on the information output using mathematical formulation and simulations. We identify key intrinsic properties of any system of interest with practical implications for sort-seq assays, depending on the experimental goals. The fluorescence range and cell-to-cell variability specify the number of sorted populations needed for quantitative measurements that are precise and unbiased. These factors also indicate cases where an enrichment-based approach that uses a single sorted population can offer satisfactory results. These predications of our model are corroborated using re-analysis of published data. We explore implications of these results for quantitative modeling and library design. Sort-seq assays can be streamlined by reducing the number of sorted populations, saving considerable resources. Simple preliminary experiments can guide optimal experiment design, minimizing cost while maintaining the maximal information output and avoiding latent biases. These insights can facilitate future applications of this highly adaptable technique." @default.
• W2294418165 created "2016-06-24" @default.
• W2294418165 creator A5051039938 @default.
• W2294418165 creator A5058666992 @default.
• W2294418165 date "2016-03-09" @default.
• W2294418165 modified "2023-10-09" @default.
• W2294418165 title "Sort-seq under the hood: implications of design choices on large-scale characterization of sequence-function relations" @default.
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• W2294418165 doi "https://doi.org/10.1186/s12864-016-2533-5" @default.
• W2294418165 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4784318" @default.
• W2294418165 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/26956374" @default.
• W2294418165 hasPublicationYear "2016" @default.
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