Polyelectrolyte Surface Interface for Single-Molecule Fluorescence Studies of DNA Polymerase
Author(s) -
Emil P. Kartalov,
Marc Unger,
Stephen R. Quake
Publication year - 2003
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/03343st02
Subject(s) - dna , dna polymerase , polymerase , biophysics , dna clamp , molecule , nucleotide , chemistry , fluorescence , polyelectrolyte , biochemistry , polymerase chain reaction , microbiology and biotechnology , biology , polymer , gene , physics , organic chemistry , reverse transcriptase , quantum mechanics
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.
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