Probing DNA-DNA Interactions with a Combination of Quadruple-Trap Optical Tweezers and Microfluidics

Research output: Chapter in Book / Report / Conference proceedingChapterAcademicpeer-review

Abstract

DNA metabolism and DNA compaction in vivo involve frequent interactions of remote DNA segments, mediated by proteins. In order to gain insight into such interactions, quadruple-trap optical tweezers have been developed. This technique provides an unprecedented degree of control through the ability to independently manipulate two DNA molecules in three dimensions. In this way, discrete regions of different DNA molecules can be brought into contact with one another, with a well-defined spatial configuration. At the same time, the tension and extension of the DNA molecules can be monitored. Furthermore, combining quadruple-trap optical tweezers with microfluidics makes fast buffer exchange possible, which is important for in situ generation of the dual DNA-protein constructs needed for these kinds of experiments. In this way, processes such as protein-mediated inter-DNA bridging can be studied with unprecedented control. This chapter provides a step-by-step description of how to perform a dual DNA manipulation experiment using combined quadruple-trap optical tweezers and microfluidics.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
EditorsJohn M. Walker
PublisherHumana Press, Methods Mol Biol
Pages275-293
Number of pages19
Volume1486
DOIs
Publication statusPublished - 2017

Keywords

  • DNA
  • DNA Probes
  • Journal Article
  • Microfluidics
  • Nucleic Acid Hybridization
  • Optical Tweezers
  • Optics and Photonics

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