Protocol for effective surface passivation for single-molecule studies of chromatin and topoisomerase II
Summary: 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...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724006658 |
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| author | Tung T. Le Xiang Gao Seong Ha Park Jaeyoon Lee James T. Inman Michelle D. Wang |
| author_facet | Tung T. Le Xiang Gao Seong Ha Park Jaeyoon Lee James T. Inman Michelle D. Wang |
| author_sort | Tung T. Le |
| collection | DOAJ |
| description | Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-e138d0de2657403b82c465ed1e3b15b0 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-e138d0de2657403b82c465ed1e3b15b02024-12-18T08:52:08ZengElsevierSTAR Protocols2666-16672025-03-0161103500Protocol for effective surface passivation for single-molecule studies of chromatin and topoisomerase IITung T. Le0Xiang Gao1Seong Ha Park2Jaeyoon Lee3James T. Inman4Michelle D. Wang5Howard Hughes Medical Institute, Cornell University, Ithaca, NY 14853, USA; Physics Department & LASSP, Cornell University, Ithaca, NY 14853, USAHoward Hughes Medical Institute, Cornell University, Ithaca, NY 14853, USA; Physics Department & LASSP, Cornell University, Ithaca, NY 14853, USABiophysics Program, Cornell University, Ithaca, NY 14853, USAPhysics Department & LASSP, Cornell University, Ithaca, NY 14853, USAHoward Hughes Medical Institute, Cornell University, Ithaca, NY 14853, USA; Physics Department & LASSP, Cornell University, Ithaca, NY 14853, USAHoward Hughes Medical Institute, Cornell University, Ithaca, NY 14853, USA; Physics Department & LASSP, Cornell University, Ithaca, NY 14853, USA; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724006658biophysicssingle-molecule assaysmolecular biology |
| spellingShingle | Tung T. Le Xiang Gao Seong Ha Park Jaeyoon Lee James T. Inman Michelle D. Wang Protocol for effective surface passivation for single-molecule studies of chromatin and topoisomerase II STAR Protocols biophysics single-molecule assays molecular biology |
| title | Protocol for effective surface passivation for single-molecule studies of chromatin and topoisomerase II |
| title_full | Protocol for effective surface passivation for single-molecule studies of chromatin and topoisomerase II |
| title_fullStr | Protocol for effective surface passivation for single-molecule studies of chromatin and topoisomerase II |
| title_full_unstemmed | Protocol for effective surface passivation for single-molecule studies of chromatin and topoisomerase II |
| title_short | Protocol for effective surface passivation for single-molecule studies of chromatin and topoisomerase II |
| title_sort | protocol for effective surface passivation for single molecule studies of chromatin and topoisomerase ii |
| topic | biophysics single-molecule assays molecular biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724006658 |
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