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PMID: 12661156 Published · ppublish English Evaluation Study Research Support, U.S. Gov't, P.H.S. Technical Report

Polyelectrolyte surface interface for single-molecule fluorescence studies of DNA polymerase.

BioTechniques ·Vol. 34 ·No. 3 ·2003-03-00 ·Pages 505-10

Kartalov EP, Unger MA, Quake SR

Abstract

We report the use of polyelectrolyte multilayers in a stable robust surface chemistry for specific anchoring of DNA to glass. The nonspecific binding of fluorescently tagged nucleotides is suppressed down to the single-molecule level, and DNA polymerase is active on the anchored DNA template. This surface-chemistry platform can be used for single-molecule studies of DNA and DNA polymerase and may be more broadly applicable for other situations in which it is important to have specific biomolecular surface chemistry with extremely low nonspecific binding.

MeSH Terms
Biotinylation/methods Coated Materials, Biocompatible/chemical synthesis,chemistry DNA/chemistry DNA-Directed DNA Polymerase/chemistry Electrolytes/chemistry Enzymes, Immobilized Glass Microscopy, Fluorescence/methods Protein Binding Substrate Specificity Surface Properties
Chemicals
Coated Materials, Biocompatible Electrolytes Enzymes, Immobilized DNA DNA-Directed DNA Polymerase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kartalov Emil P
Department of Applied Physics, MC128-95, Caltech, Pasadena, CA 91125, USA.
Unger Marc A
Quake Stephen R
Article Info
Journal
BioTechniques
Abbr.
Biotechniques
ISSN
0736-6205
Published
2003-03-00
Pages
505-10
Language
English
Region
England
NLM ID
8306785
Subset
IM
Grants
NIGMS NIH HHS · 5T32GM07616 · United States
NHGRI NIH HHS · HG01642 · United States
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