A simple DNA handle attachment method for single molecule mechanical manipulation experiments

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 o...

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Veröffentlicht in:Protein science 2016-08, Vol.25 (8), p.1535-1544
Hauptverfasser: Min, Duyoung, Arbing, Mark A., Jefferson, Robert E., Bowie, James U.
Format: Artikel
Sprache:eng
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Zusammenfassung: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.
ISSN:0961-8368
1469-896X
DOI:10.1002/pro.2952