Rxivist logo

An Automated Bayesian Pipeline for Rapid Analysis of Single-Molecule Binding Data

By Carlas S. Smith, Karina Jouravleva, Maximiliaan Huisman, Samson M Jolly, Phillip D. Zamore, David Grunwald

Posted 11 Apr 2018
bioRxiv DOI: 10.1101/261917 (published DOI: 10.1038/s41467-018-08045-5)

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.

Download data

  • Downloaded 532 times
  • Download rankings, all-time:
    • Site-wide: 40,239 out of 118,180
    • In molecular biology: 1,172 out of 3,617
  • Year to date:
    • Site-wide: 98,589 out of 118,180
  • Since beginning of last month:
    • Site-wide: 103,071 out of 118,180

Altmetric data

Downloads over time

Distribution of downloads per paper, site-wide


Sign up for the Rxivist weekly newsletter! (Click here for more details.)