Основная статья http://www.sciencemag.org/cgi/content/abstract/299/5607/682Science 31 January 2003: Vol. 299. no. 5607, pp. 682 - 686 DOI: 10.1126/science.1079700 REPORTSZero-Mode Waveguides for Single-Molecule Analysis at High ConcentrationsM. J. Levene,1 J. Korlach,12 S. W. Turner,1* M. Foquet,1 H. G. Craighead,1 W. W. Webb1 Optical approaches for observing the dynamics of single molecules have required pico- to nanomolar concentrations of fluorophore in order to isolate individual molecules. However, many biologically relevant processes occur at micromolar ligand concentrations, necessitating a reduction in the conventional observation volume by three orders of magnitude. We show that arrays of zero-mode waveguides consisting of subwavelength holes in a metal film provide a simple and highly parallel means for studying single-molecule dynamics at micromolar concentrations with microsecond temporal resolution. We present observations of DNA polymerase activity as an example of the effectiveness of zero-mode waveguides for performing single-molecule experiments at high concentrations. 1 Applied and Engineering Physics, 2 Graduate Program in Biochemistry, Molecular, and Cell Biology, Cornell University, Clark Hall, Ithaca, NY 14853, USA. * Present address: Nanofluidics Incorporated, 17 Sheraton Drive, Ithaca, NY 14850, USA.
To whom correspondence should be addressed. E-mail: www2@cornell.edu
Read the Full Text
The editors suggest the following Related Resources on Science sites:In Science Magazine- PERSPECTIVES
ANALYTICAL CHEMISTRY: How to Detect Weak Pairs - Ted A. Laurence and Shimon Weiss (31 January 2003)
Science 299 (5607), 667. [DOI: 10.1126/science.1081025] Summary » Full Text » PDF »
THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:- From the Cover: Single-nucleotide discrimination in immobilized DNA oligonucleotides with a biological nanopore.
- D. Stoddart, A. J. Heron, E. Mikhailova, G. Maglia, and H. Bayley (2009)
PNAS 106, 7702-7707 Abstract » Full Text » PDF »
- Mutants of Taq DNA polymerase resistant to PCR inhibitors allow DNA amplification from whole blood and crude soil samples.
- M. B. Kermekchiev, L. I. Kirilova, E. E. Vail, and W. M. Barnes (2009)
Nucleic Acids Res. 37, e40 Abstract » Full Text » PDF »
- Next-Generation Sequencing: From Basic Research to Diagnostics.
- K. V. Voelkerding, S. A. Dames, and J. D. Durtschi (2009)
Clin. Chem. 55, 641-658 Abstract » Full Text » PDF »
- Real-Time DNA Sequencing from Single Polymerase Molecules.
- J. Eid, A. Fehr, J. Gray, K. Luong, J. Lyle, G. Otto, P. Peluso, D. Rank, P. Baybayan, B. Bettman, et al. (2009)
Science 323, 133-138 Abstract » Full Text » PDF »
- Keeping Up With the Next Generation: Massively Parallel Sequencing in Clinical Diagnostics.
- J. R. ten Bosch and W. W. Grody (2008)
J. Mol. Diagn. 10, 484-492 Abstract » Full Text » PDF »
- An artificial processivity clamp made with streptavidin facilitates oriented attachment of polymerase-DNA complexes to surfaces.
- J. G. K. Williams, D. L. Steffens, J. P. Anderson, T. M. Urlacher, D. T. Lamb, D. L. Grone, and J. C. Egelhoff (2008)
Nucleic Acids Res. 36, e121 Abstract » Full Text » PDF »
- Effect of the C-terminal Truncation on the Functional Cycle of Chaperonin GroEL: IMPLICATION THAT THE C-TERMINAL REGION FACILITATES THE TRANSITION FROM THE FOLDING-ARRESTED TO THE FOLDING-COMPETENT STATE.
- M. Suzuki, T. Ueno, R. Iizuka, T. Miura, T. Zako, R. Akahori, T. Miyake, N. Shimamoto, M. Aoki, T. Tanii, et al. (2008)
J. Biol. Chem. 283, 23931-23939 Abstract » Full Text » PDF »
- Selective aluminum passivation for targeted immobilization of single DNA polymerase molecules in zero-mode waveguide nanostructures.
- J. Korlach, P. J. Marks, R. L. Cicero, J. J. Gray, D. L. Murphy, D. B. Roitman, T. T. Pham, G. A. Otto, M. Foquet, and S. W. Turner (2008)
PNAS 105, 1176-1181 Abstract » Full Text » PDF »
- Single-molecule biophysics: at the interface of biology, physics and chemistry.
- A. A Deniz, S. Mukhopadhyay, and E. A Lemke (2008)
J R Soc Interface 5, 15-45 Abstract » Full Text » PDF »
- Genome-wide analysis of DNA methylation patterns.
- D. Zilberman and S. Henikoff (2007)
Development 134, 3959-3965 Abstract » Full Text » PDF »
- Single-molecule Chemistry and Biology Special Feature: Fueling protein DNA interactions inside porous nanocontainers.
- I. Cisse, B. Okumus, C. Joo, and T. Ha (2007)
PNAS 104, 12646-12650 Abstract » Full Text » PDF »
- Measuring Rotations of a Few Cross-Bridges in Skeletal Muscle.
- J. Borejdo, J. Talent, and I. Akopova (2006)
Exp Biol Med 231, 28-38 Abstract » Full Text » PDF »
- Emerging technologies in DNA sequencing.
- M. L. Metzker (2005)
Genome Res. 15, 1767-1776 Abstract » Full Text » PDF »
- Temporally resolved interactions between antigen-stimulated IgE receptors and Lyn kinase on living cells.
- D. R. Larson, J. A. Gosse, D. A. Holowka, B. A. Baird, and W. W. Webb (2005)
J. Cell Biol. 171, 527-536 Abstract » Full Text » PDF »
- Simple, robust methods for high-throughput nanoliter-scale DNA sequencing.
- D. E. Smailus, A. Marziali, P. Dextras, M. A. Marra, and R. A. Holt (2005)
Genome Res. 15, 1447-1450 Abstract » Full Text » PDF »
- Accurate Multiplex Polony Sequencing of an Evolved Bacterial Genome.
- J. Shendure, G. J. Porreca, N. B. Reppas, X. Lin, J. P. McCutcheon, A. M. Rosenbaum, M. D. Wang, K. Zhang, R. D. Mitra, and G. M. Church (2005)
Science 309, 1728-1732 Abstract » Full Text » PDF »
- Surface plasmon field-enhanced fluorescence spectroscopy studies of primer extension reactions.
- G. Stengel and W. Knoll (2005)
Nucleic Acids Res. 33, e69 Abstract » Full Text » PDF »
- Fluorescence-aided molecule sorting: Analysis of structure and interactions by alternating-laser excitation of single molecules.
- A. N. Kapanidis, N. K. Lee, T. A. Laurence, S. Doose, E. Margeat, and S. Weiss (2004)
PNAS 101, 8936-8941 Abstract » Full Text » PDF »
- Enzyme screening with synthetic multifunctional pores: Focus on biopolymers.
- N. Sorde, G. Das, and S. Matile (2003)
PNAS 100, 11964-11969 Abstract » Full Text » PDF »
- The structural basis for giant enhancement enabling single-molecule Raman scattering.
- Z. Wang, S. Pan, T. D. Krauss, H. Du, and L. J. Rothberg (2003)
PNAS 100, 8638-8643 Abstract » Full Text » PDF »
|