An Automated Bayesian Pipeline for Rapid Analysis of Single-Molecule Binding Data [preprint]
Authors
Smith, CarlasJouravleva, Karina
Huisman, Maximiliaan
Jolly, Samson M.
Zamore, Phillip D.
Grunwald, David
UMass Chan Affiliations
RNA Therapeutics InstituteDocument Type
PreprintPublication Date
2018-04-11Keywords
Automated Bayesian PipelineSingle-Molecule Binding Data
TtAgo
Thermus thermophilus
DNA
Molecular Biology
Genetic Phenomena
Molecular Biology
Metadata
Show full item recordAbstract
Single-molecule binding assays enable the study of how molecular machines assemble and function. Current algorithms can identify and locate individual molecules, but require tedious manual validation of each spot. Moreover, no solution for high-throughput analysis of single-molecule binding data exists. Here, we describe an automated pipeline to analyze single-molecule data over a wide range of experimental conditions. We benchmarked the pipeline by measuring the binding properties of the well-studied, DNA-guided DNA endonuclease, TtAgo, an Argonaute protein from the Eubacterium Thermus thermophilus. We also used the pipeline to extend our understanding of TtAgo by measuring the protein's binding kinetics at physiological temperatures and for target DNAs containing multiple, adjacent binding sites.Source
bioRxiv 261917; doi: https://doi.org/10.1101/26191. Link to preprint on bioRxiv service.
DOI
10.1101/261917Permanent Link to this Item
http://hdl.handle.net/20.500.14038/29268Rights
The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.Distribution License
http://creativecommons.org/licenses/by-nc-nd/4.0/ae974a485f413a2113503eed53cd6c53
10.1101/261917
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Except where otherwise noted, this item's license is described as The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.