Protocol for effective surface passivation for single-molecule studies of chromatin and topoisomerase II.

Autor: Le TT; Howard Hughes Medical Institute, Cornell University, Ithaca, NY 14853, USA; Physics Department & LASSP, Cornell University, Ithaca, NY 14853, USA., Gao X; Howard Hughes Medical Institute, Cornell University, Ithaca, NY 14853, USA; Physics Department & LASSP, Cornell University, Ithaca, NY 14853, USA., Ha Park S; Biophysics Program, Cornell University, Ithaca, NY 14853, USA., Lee J; Physics Department & LASSP, Cornell University, Ithaca, NY 14853, USA., Inman JT; Howard Hughes Medical Institute, Cornell University, Ithaca, NY 14853, USA; Physics Department & LASSP, Cornell University, Ithaca, NY 14853, USA., Wang MD; Howard Hughes Medical Institute, Cornell University, Ithaca, NY 14853, USA; Physics Department & LASSP, Cornell University, Ithaca, NY 14853, USA. Electronic address: mwang@physics.cornell.edu.
Jazyk: angličtina
Zdroj: STAR protocols [STAR Protoc] 2024 Dec 17; Vol. 6 (1), pp. 103500. Date of Electronic Publication: 2024 Dec 17.
DOI: 10.1016/j.xpro.2024.103500
Abstrakt: For single-molecule studies requiring surface anchoring of biomolecules, poorly passivated surfaces can result in alterations of biomolecule structure and function that lead to artifacts. Here, we present a surface passivation assay for single-molecule studies of chromatin and topoisomerase II. We detail steps for preparing a nucleosome array and hydrophobic nitrocellulose-coated flow cell. We then describe procedures for chromatin stretching with an angular optical trap (AOT) and performing a chromatin-topoisomerase experiment. This method is cost effective and potentially applicable to other biomolecules. For complete details on the use and execution of this protocol, please refer to Le et al. 1 .
Competing Interests: Declaration of interests The authors declare no competing interests.
(Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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