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A simple DNA handle attachment method for single molecule mechanical manipulation experiments
Author(s) -
Min Duyoung,
Arbing Mark A.,
Jefferson Robert E.,
Bowie James U.
Publication year - 2016
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.2952
Subject(s) - dna , molecule , biological system , tethering , nanotechnology , covalent bond , molecular biophysics , biophysics , reagent , chemistry , computer science , materials science , biology , biochemistry , organic chemistry , operating system
Manipulating single molecules and systems of molecules with mechanical force is a powerful technique to examine their physical properties. Applying force requires attachment of the target molecule to larger objects using some sort of molecular tether, such as a strand of DNA. DNA handle attachment often requires difficult manipulations of the target molecule, which can preclude attachment to unstable, hard to obtain, and/or large, complex targets. Here we describe a method for covalent DNA handle attachment to proteins that simply requires the addition of a preprepared reagent to the protein and a short incubation. The handle attachment method developed here provides a facile approach for studying the biomechanics of biological systems.