A versatile method for generating single DNA molecule patterns: Through the combination of directed capillary assembly a
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Jérôme Chalmeau Physics Department, University of Minnesota, Minneapolis, Minnesota 55416; CNRS, LAAS, F-31077 Toulouse, France; and Université de Toulouse, UPS, INSA, INP, ISAE, LAAS, F-31077 Toulouse, France
Angélique Coutable Université de Toulouse, INSA, UPS, INP, LISBP, Toulouse F-31077, France; and INRA, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, CNRS, UMR5504, Toulouse F-31400, France
Christophe Vieu CNRS, LAAS, F-31077 Toulouse, France; and Université de Toulouse, UPS, INSA, INP, ISAE, LAAS, F-31077 Toulouse, France (Received 21 May 2010; accepted 4 August 2010)
One of the challenges in the development of molecular scale devices is the integration of nano-objects or molecules onto desired locations on a surface. This integration comprises their accurate positioning, their alignment, and the preservation of their functionality. Here, we proved how capillary assembly in combination with soft lithography can be used to perform DNA molecular combing to generate chips of isolated DNA strands for genetic analysis and diagnosis. The assembly of DNA molecules is achieved on a topologically micropatterned polydimethylsiloxane stamp inducing almost simultaneously the trapping and stretching of single molecules. The DNA molecules are then transferred onto aminopropyltriethoxysilane-coated surfaces. In fact, this technique offers the possibility to tightly control the experimental parameters to direct the assembly process. This technique does not induce a selection in size of the objects, therefore it can handle complex solutions of long (tens of kbp) but also shorter (a few thousands of bp) molecules directly in solution to allow the construction of future one-dimensional nanoscale building templates.
Address all correspondence to this author. e-mail: [email protected] DOI: 10.1557/jmr.2010.12
mapping5 and medical diagnostics.6 However, the current tendency is to be able to pattern individual stretched DNA molecules into a well-defined array so that a fast optical readout can be performed. The generation of such a controlled assembly could be a major building block for the construction of future devices. In fact, DNA chains can be metalized7–11 or functionalized with nanoparticles12 and this ability to generate well-aligned one-dimensional (1D) nanoscale templates onto desired locations on a solid support could provide further opportunities for the fabrication of assemblies with increasing complexity. In the literature, a number of studies in molecular combing13–15 have been reported. The so-called “molecular combing” technique was introduced in 1993, and performed on short DNA molecules. This technique consists in obtaining from a coiled DNA molecule in its natural state, aligned and stretched DNA filaments that are easier to study than under their compacted state. For this purpose, a glass support chemically treated is immersed into a solution containing the DNA molecules. Each molecule gets fixed by one of its extremities. The glass
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Ó Materials Research Society 2011
I. INTRODUCTION
The fabrica
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