Single molecule studies of helicases with magnetic tweezers.

Publication Type:

Journal Article

Source:

Methods (San Diego, Calif.), Volume 105, p.3-15 (2016)

DOI:

10.1016/j.ymeth.2016.06.019

Keywords:

DNA; DNA Helicases; DNA Replication; DNA, Cruciform; Magnetics; OPTICAL TWEEZERS; RNA; Single Molecule Imaging

Abstract:

Helicases are a broad family of enzymes that perform crucial functions in DNA replication and in the maintenance of DNA and RNA integrity. A detailed mechanical study of helicases on DNA and RNA is possible using single molecule manipulation methods. Among those, magnetic tweezers (or traps) present a convenient, moderate throughput assay (tens of enzymes can be monitored simultaneously) that allow for high resolution (single base-pair) studies of these enzymes in various conditions and on various substrates (double and single stranded DNA and RNA). Here we discuss various implementation of the basic assay relevant for these studies.