Simple method of DNA stretching on glass substrate for fluorescence imaging and spectroscopy

Abstract. We demonstrate a simple method of stretching DNA to its full length, suitable for optical imaging and atomic force microscopy (AFM). Two competing forces on the DNA molecules, which are the electrostatic attraction between positively charged dye molecules (YOYO-1) intercalated into DNA and the negatively charged surface of glass substrate, and the centrifugal force of the rotating substrate, are mainly responsible for the effective stretching and the dispersion of single strands of DNA. The density of stretched DNA molecules could be controlled by the concentration of the dye-stained DNA solution. Stretching of single DNA molecules was confirmed by AFM imaging and the photoluminescence spectra of single DNA molecule stained with YOYO-1 were obtained, suggesting that our method is useful for spectroscopic analysis of DNA at the single molecule level.

[1]  A. Bensimon,et al.  Encephalitozoon cuniculi (Microspora) genome: physical map and evidence for telomere-associated rDNA units on all chromosomes. , 2000, Nucleic acids research.

[2]  Jong-Dal Hong,et al.  Layer-by-Layer Deposited Multilayer Assemblies of Ionene-Type Polyelectrolytes Based on the Spin-Coating Method , 2001 .

[3]  Vlastimil Masek,et al.  Raman spectroscopy of DNA modified by intrastrand cross-links of antitumor cisplatin. , 2007, Journal of structural biology.

[4]  H. Hansma,et al.  Atomic force microscopy of long and short double-stranded, single-stranded and triple-stranded nucleic acids. , 1996, Nucleic acids research.

[5]  C. Zhu,et al.  A convenient method of aligning large DNA molecules on bare mica surfaces for atomic force microscopy. , 1998, Nucleic acids research.

[6]  Honggu Chun,et al.  Multi-order dynamic range DNA sensor using a gold decorated SWCNT random network. , 2011, ACS nano.

[7]  Daniel J. Müller,et al.  Atomic force microscopy: A forceful way with single molecules , 1999, Current Biology.

[8]  F. Crick,et al.  Molecular Structure of Nucleic Acids: A Structure for Deoxyribose Nucleic Acid , 1953, Nature.

[9]  A Bensimon,et al.  Alignment and sensitive detection of DNA by a moving interface. , 1994, Science.

[10]  D. Klinov,et al.  Observation of single‐stranded DNA on mica and highly oriented pyrolytic graphite by atomic force microscopy , 2006, FEBS letters.

[11]  S. Smith,et al.  Direct mechanical measurements of the elasticity of single DNA molecules by using magnetic beads. , 1992, Science.

[12]  Kookheon Char,et al.  Fabrication of Highly Ordered Multilayer Films Using a Spin Self‐Assembly Method , 2001 .

[13]  Hyun Jun Kim,et al.  Confocal Raman spectroscopy of single poly(3-methylthiophene) nanotubes , 2007 .

[14]  Philip LeDuc,et al.  Dynamics of individual flexible polymers in a shear flow , 1999, Nature.

[15]  H. W. Liu,et al.  Materials science communication effects of rca clean-up procedures on the formation of roughened poly-si electrodes for high-density drams' capacitors , 1997 .

[16]  R. Fraser The structure of deoxyribose nucleic acid. , 2004, Journal of structural biology.

[17]  B. Finlayson‐Pitts,et al.  Substrate changes associated with the chemistry of self-assembled monolayers on silicon. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[18]  B. Berge,et al.  Observation by fluorescence microscopy of transcription on single combed DNA , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[19]  N. Uyeda,et al.  Direct imaging of a double-strand DNA molecule. , 1981, Ultramicroscopy.

[20]  M. Castillejo,et al.  Gold coating of micromechanical DNA biosensors by pulsed laser deposition , 2012 .

[21]  Michelle D. Wang,et al.  Stretching of single collapsed DNA molecules. , 2000, Biophysical journal.

[22]  D E Wemmer,et al.  Stable fluorescent complexes of double-stranded DNA with bis-intercalating asymmetric cyanine dyes: properties and applications. , 1992, Nucleic acids research.

[23]  A. M. Brett,et al.  Atomic Force Microscopy of DNA Immobilized onto a Highly Oriented Pyrolytic Graphite Electrode Surface , 2003 .

[24]  D. Pang,et al.  Combing DNA on CTAB-coated surfaces. , 2004, Biophysical chemistry.

[25]  M. Muramatsu,et al.  J. D. Watson & F. H. C. Crick: A Structure for Deoxyribose Nucleic Acid , 2002 .

[26]  S Povey,et al.  Dynamic molecular combing: stretching the whole human genome for high-resolution studies. , 1997, Science.

[27]  Olivier Hyrien,et al.  A simple and optimized method of producing silanized surfaces for FISH and replication mapping on combed DNA fibers. , 2008, BioTechniques.

[28]  Olle Inganäs,et al.  Soft lithographic printing of patterns of stretched DNA and DNA/electronic polymer wires by surface-energy modification and transfer. , 2006, Small.

[29]  X. Michalet,et al.  High-resolution mapping of the X-linked lymphoproliferative syndrome region by FISH on combed DNA , 1998, Cytogenetic and Genome Research.

[30]  W. Li,et al.  Surface-engineered gold nanorods: promising DNA vaccine adjuvant for HIV-1 treatment. , 2012, Nano letters.