Journal
STRUCTURE
Volume 28, Issue 12, Pages 1337-+Publisher
CELL PRESS
DOI: 10.1016/j.str.2020.07.009
Keywords
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Funding
- NIBIB grant [EB-002027]
- National Institutes of Health [5P01CA112181-05]
- DARPA [W911NF-06-1-0122]
- NSF [0647161]
- NASA [NNX07AK54G]
- Louis V. Gerstner, Jr. Young Investigators Fund
- National Cancer Institute grant [P30 CA008748]
- National Institutes of Health (NIH) Director's New Innovator Award [1DP2GM105443-01]
- BBSRC [BB/L021730/1] Funding Source: UKRI
- Direct For Mathematical & Physical Scien
- Division Of Physics [0647161] Funding Source: National Science Foundation
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Single-molecule detection and manipulation is a powerful tool for unraveling dynamic biological processes. Unfortunately, success in such experiments is often challenged by tethering the biomolecule(s) of interest to a biocompatible surface. Here, we describe a robust surface passivation method by dense polymer brush grafting, based on optimized polyethylene glycol (PEG) deposition conditions, exactly at the lower critical point of an aqueous biphasic PEG-salt system. The increased biocompatibility achieved, compared with PEG deposition in sub-optimal conditions away from the critical point, allowed us to successfully detect the assembly and function of a large macromolecular machine, a fluorescent-labeled multi-subunit, human RNA Polymerase II Transcription Pre-Initiation Complex, on single, promoter-containing, surface-immobilized DNA molecules. This platform will enable probing the complex biochemistry and dynamics of large, multi-subunit macromolecular assemblies, such as during the initiation of human RNA Pol II transcription, at the single-molecule level.
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