Single molecule measurements of DNA helicase activity with magnetic tweezers and t-test based step-finding analysis.

ABSTRACT: Magnetic tweezers is a versatile and easy to implement single-molecule technique that has become increasingly prevalent in the study of nucleic acid based molecular motors. Here, we provide a description of the magnetic tweezers instrument and guidelines for measuring and analyzing DNA helicase activity. Along with experimental methods, we describe a robust method of single-molecule trajectory analysis based on the Student's t-test that accommodates continuous transitions in addition to the discrete transitions assumed in most widely employed analysis routines. To illustrate the single-molecule unwinding assay and the analysis routine, we provide DNA unwinding measurements of Escherichia coli RecQ helicase under a variety of conditions (Na+, ATP, temperature, and DNA substrate geometry). T

SUBMITTER: Seol Y 

PROVIDER: S-EPMC4967025 | biostudies-literature | 2016 Aug

REPOSITORIES: biostudies-literature