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

Biotechniques. 2003 Mar;34(3):505-10. doi: 10.2144/03343st02.

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.

Publication types

  • Evaluation Study
  • Research Support, U.S. Gov't, P.H.S.
  • Technical Report

MeSH terms

  • Biotinylation / methods
  • Coated Materials, Biocompatible / chemical synthesis*
  • Coated Materials, Biocompatible / chemistry
  • DNA / chemistry*
  • DNA-Directed DNA Polymerase / chemistry*
  • Electrolytes / chemistry*
  • Enzymes, Immobilized
  • Glass
  • Microscopy, Fluorescence / methods*
  • Protein Binding
  • Substrate Specificity
  • Surface Properties

Substances

  • Coated Materials, Biocompatible
  • Electrolytes
  • Enzymes, Immobilized
  • DNA
  • DNA-Directed DNA Polymerase