An Effective Surface Passivation Assay for Single-Molecule Studies of Chromatin and Topoisomerase II

一种用于染色质和拓扑异构酶 II 单分子研究的有效表面钝化试验

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作者:Tung T Le, Xiang Gao, Seong Ha Park, Jaeyoon Lee, James T Inman, Michelle D Wang

Abstract

For complete details on the use and execution of this protocol, please refer to Le et al. (2019)1. Summary: A.For single-molecule studies requiring surface anchoring of biomolecules, a poorly passivated surface can result in alterations of biomolecule structure and function that can result in artifacts. This protocol describes surface passivation and sample chamber preparation for mechanical manipulation of chromatin fibers and characterization of topoisomerase II activity in physiological buffer conditions. The method employs enhanced surface hydrophobicity and purified blocking proteins to reduce non-specific surface adsorption. This method is accessible, cost-effective, and potentially widely applicable to other biomolecules.For a complete list of publications that employ this protocol, see the paper references.

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