Tethered Particle Motion: An Easy Technique for Probing DNA Topology and Interactions with Transcription Factors.

ABSTRACT: Tethered Particle Motion (TPM) is a versatile in vitro technique for monitoring the conformations a linear macromolecule, such as DNA, can exhibit. The technique involves monitoring the diffusive motion of a particle anchored to a fixed point via the macromolecule of interest, which acts as a tether. In this chapter, we provide an overview of TPM, review the fundamental principles that determine the accuracy with which effective tether lengths can be used to distinguish different tether conformations, present software tools that assist in capturing and analyzing TPM data, and provide a protocol which uses TPM to characterize lac repressor-induced DNA looping. Critical to any TPM assay is the understanding of the timescale over which the diffusive motion of the particle must be observed to

SUBMITTER: Kovari DT 

PROVIDER: S-EPMC6089228 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature