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Modeling Nanopores for Sequencing DNA

Using nanopores to sequence DNA rapidly and at a low cost has the potential to radically transform the field of genomic research. However, despite all the exciting developments in the field, sequencing DNA using a nanopore has yet to be demonstrated. Among the many problems that hinder developme ...

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Measuring DNAProtein Binding Affinity on a Single Molecule Using Optical Tweezers

DNA–protein interactions may be observed on single molecules with a variety of techniques. However, quantifying the binding affinity is difficult and often requires many (∼100) individual events to characterize the interaction. We use a single λ DNA molecule that provides a lattice of bin ...

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Engineering Mononucleosomes for Single-Pair FRET Experiments

In DNA nanotechnology, DNA is used as a structural material, rather than as an information carrier. The structural organization of the DNA itself determines accessibility to its underlying information content in vivo. Nucleosomes form the basic level of DNA compaction in eukaryotic nuc ...

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Synthesis and Characterization of Self-Assembled DNA Nanostructures

The past decade witnessed the fast evolvement of structural DNA nanotechnology, which uses DNA as blueprint and building material to construct artificial nanostructures. Using branched DNA as the main building block (also known as a “tile”) and cohesive single-stranded DNA (ssDNA) ends ...

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Optimal Practices for Surface-Tethered Single Molecule Total Internal Reflection Fluorescence Resonance Energy Transfer Analysis

Single molecule fluorescence microscopy can be used to follow the mechanics of molecular biology processes in real time. However, many factors, from flow cell preparation to improper data analysis can negatively impact single molecule fluorescence resonance energy transfer (sm ...

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DNA Molecular Handles for Single-Molecule Protein-Folding Studies by Optical Tweezers

In this chapter, we describe a method that extends the use of optical tweezers to the study of the folding mechanism of single protein molecules. This method entails the use of DNA molecules as molecular handles to manipulate individual proteins between two polystyrene beads. The DNA molecules ...

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DNA Contour Length Measurements as a Tool for the Structural Analysis of DNA and Nucleoprotein Complexes

The atomic force microscope (AFM) is a widely used tool to image DNA and nucleoprotein complexes at the molecular level. This is because the AFM is relatively easy to operate, has the capability to image biomolecules under aqueous solutions, and, most importantly, can image mesoscopic macromo ...

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Trapping and Immobilization of DNA Molecules Between Nanoelectrodes

DNA is one of the most promising molecules for nanoscale bottom-up fabrication. For both scientific studies and fabrication of devices, it is desirable to be able to manipulate DNA molecules, or self-�assembled DNA constructions, at the single unit level. Efficient methods are needed for pre ...

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The Atomic Force Microscopy as a Lithographic Tool: Nanografting of DNA Nanostructures for Biosensing Applications

Current in vitro techniques cannot accurately identify small differences in concentration in samples containing few molecules in single or few cells. Nanotechnology overcomes these limitations with the possibility of measuring protein amounts down to a hundred molecules and s ...

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DNA-Modified Single Crystal and Nanoporous Silicon

The functionalization of silicon as elemental crystalline wafer, nanoporous layers, or nanocrystalline particles with DNA oligonucleotides using automated solid phase synthesis is described. The procedures provide semiconductor surfaces covalently modified with o ...

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Templated Assembly of DNA Origami Gold Nanoparticle Arrays on Lithographically Patterned Surfaces

Artificial DNA nanostructures such as DNA origami have garnered significant interest as templates for sub-20 nm lithography because their rational design allows for the incorporation of binding sites to assemble nanocomponents with 6 nm resolution. In addition, their overall size ...

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Controlled Confinement of DNA at the Nanoscale: Nanofabrication and Surface Bio-Functionalization

Nanopatterned arrays of biomolecules are a powerful tool to address fundamental issues in many areas of biology. DNA nanoarrays, in particular, are of interest in the study of DNA–protein interactions and for biodiagnostic investigations. In this context, achieving a highly specific n ...

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Preparation of DNA Nanostructures with Repetitive Binding Motifs by Rolling Circle Amplification

A long one-dimensional single-stranded DNA (ssDNA) molecule with a periodic sequence motif is an attractive building block for DNA nanotechnology because it allows the positioning of oligonucleotide-labeled particles or molecules with high spatial resolution via molecular s ...

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G-Wire Synthesis and Modification with Gold Nanoparticle

DNA molecules are well known for containing the genetic information of an individual. Furthermore, DNA is a biopolymer with the potential of building up nanoscale structures. These structures can be addressed sequence specifically and, therefore, they allow connecting and arrangi ...

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A TiO2 Nanoparticle System for Sacrificial Solar H2 Production Prepared by Rational Combination of a Hydrogenase with a Ruthenium Photosensitizer

A hybrid system comprising a hydrogenase and a photosensitizer co-attached to a nanoparticle serves as a rational model for fast dihydrogen (H2) production using visible light. This chapter describes a stepwise procedure for preparing TiO2 nanoparticles functionalized with a hyd ...

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Reversible His-Tagged Enzyme Immobilization on Functionalized Carbon Nanotubes as Nanoscale Biocatalyst

Common enzyme immobilization methods on nanomaterials (adsorption, covalent binding, crosslinking, encapsulation) often generate problems in enzyme leaching, 3D structure change and diffusion resistance. We show here a detailed site-specific enzyme immobilization m ...

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Kinesin I ATPase Manipulates Biohybrids Formed from Tubulin and Carbon Nanotubes

This chapter describes a method for the formation of novel protein–nanotube hybrid conjugates. Specifically, we took advantage of the self-assembly and self-recognition properties of tubulin cytoskeletal protein immobilized onto carbon nanotubes to form nanotube-based bi ...

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Enzyme-Immobilized CNT Network Probe for In Vivo Neurotransmitter Detection

Glutamate is the principal excitatory neurotransmitter in the brain, and its excessive release plays a key role in neuronal death associated with a wide range of neural disorders. Real-time monitoring of extracellular glutamate levels would be very helpful in understanding the excit ...

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Organic-Soluble Enzyme Nano-Complexes Formed by Ion-Pairing with Surfactants

The solubilization of enzymes in organic solvents for non-aqueous biocatalysis has attracted considerable attention since the homogeneous distribution accounts for a drastically improved reaction efficiency compared to enzymes dispersed as aggregates in an organic pha ...

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Molecular Assembly-Assisted Biocatalytic Reactions in Ionic Liquids

Room temperature ionic liquids (RTILs), having no measurable vapor pressure, represent an interesting class of tunable designer solvents. Due to their many unique properties, ILs have been used as attractive alternatives to environmentally harmful ordinary organic solvents in a w ...

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