DNA Y structure: a versatile, multidimensional single molecule assay. Academic Article uri icon

Overview

abstract

  • Optical trapping is a powerful single molecule technique used to study dynamic biomolecular events, especially those involving DNA and DNA-binding proteins. Current implementations usually involve only one of stretching, unzipping, or twisting DNA along one dimension. To expand the capabilities of optical trapping for more complex measurements would require a multidimensional technique that combines all of these manipulations in a single experiment. Here, we report the development and utilization of such a novel optical trapping assay based on a three-branch DNA construct, termed a "Y structure". This multidimensional assay allows precise, real-time tracking of multiple configurational changes. When the Y structure template is unzipped under both force and torque, the force and extension of all three branches can be determined simultaneously. Moreover, the assay is readily compatible with fluorescence, as demonstrated by unzipping through a fluorescently labeled, paused transcription complex. This novel assay thus allows for the visualization and precision mapping of complex interactions of biomechanical events.

publication date

  • October 14, 2014

Research

keywords

  • DNA
  • Optical Tweezers

Identity

PubMed Central ID

  • PMC4245981

Scopus Document Identifier

  • 84910001921

Digital Object Identifier (DOI)

  • 10.1021/nl503009d

PubMed ID

  • 25291441

Additional Document Info

volume

  • 14

issue

  • 11