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Single Nucleotide Discrimination with Electro-Optical Nanopore
656a
Wednesday, March 2, 2016
that reaction sensitivity of the hydrogel encapsulated biosensor was increased by molecular confinement effect. Furthermore, hydrogel in a bead form was created using a droplet generating microfluidic device that has flow focusing microchannels. Reaction sensitivity was futher enhanced when biosensors were encapsulated in a bead form. In this case, phosphinothricin acetyltransferase (PAT), which is herbicide resistant protein that is related to marker of genetically modified (GM) crops was used as a model target. Using hydrogel bead biosensor presented in this study, PAT protein can be detected with naked eyes in a concentration as low as 20nM. This concept to detect low concentration of biomolecules can be potentially applied to other biosensors or analytical devices. 3239-Pos Board B616 Single Nucleotide Discrimination with Electro-Optical Nanopore Chan Cao, Yi-Tao Long. Ecust, Shanghai, China. Nanopore is an emerging technique, during the past two decades, has been explore to analysis the information about the identity, concentration, structure, and dynamics of the target molecules. However, the readout signal is mainly related on the tiny changes of ionic current as single molecules pass through the nanopore. Here, we develop the ultra-low current analyzer and combine plasmonic resonance scattering spectroscopy with nanopore technique, which could acquire the optical-electrical dual signals as a molecule pass though the nanopore. It provides more unique information of single biomolecule, which have a significant potential in life science. By virtue of this integrated strategy, we have realize single nucleotide discrimination, and even for the detecting the substituent difference in a molecule, such as DNA cytosine methylation. 1. Yi-Tao Long, Rui Gao, Chan Cao, Yi-Lun Ying, Da-Wei Li. CN203572764U[P]. 2. Gang Logan Liu, Yi-Tao Long, Yeonho Choi, Taewook Kang & Luke P Lee, Nat. Methods, 2007, 4(12), 1015-1017.
3240-Pos Board B617 Entropically Controlled Nanomechanical DNA Origami Devices Michael W. Hudoba1, Yi Luo2, Randy Patton1, Michael G. Poirier2, Carlos Castro1. 1 Mechanical & Aerospace Engineering, The Ohio State University, Columbus, OH, USA, 2Physics, The Ohio State University, Columbus, OH, USA. Structural DNA nanotechnology is a rapidly emerging field with immense potential for applications such as single molecule sensing, drug delivery, and manipulating molecular components. Major advances in the last decade have enabled the precise design and fabrication of DNA nanostructures with unprecedented geometric complexity; however, relative to natural biomolecular machines, the functional scope of DNA nanotechnology is limited by an inability to design dynamic mechanical behavior such as complex motion, conformational dynamics, or force generation. Inspired by approaches used in macroscopic machine design, we have recently developed methods to design structures with controllable 1D, 2D, or 3D motion. Moving beyond design of geometry or motion paths, we have recently developed structures with multiple thermally accessible states separated by well-defined energy barriers with tunable height. In particular, we have design a two-state device that transitions between compact and open states driven by thermal energy. We directly measured the dynamic behavior via single molecule fluorescence experiments, which show both the equilibrium distribution of states and the kinetics of switching between states can be tuned via design parameters that regulate the relative configurational space, or equivalently the entropy change, between states. Finally, we show that these dynamic structures can be used to probe local molecular scale forces, specifically depletion forces that result from entropic interactions with a crowding reagent. This work is the first demonstration of an ability to control the kinetics of DNA origami nanostructures down to the second scale and establishes a foundation for exploiting dynamic behavior of these devices to study physical interactions at the molecular scale.
Wednesday, March 2, 2016
that reaction sensitivity of the hydrogel encapsulated biosensor was increased by molecular confinement effect. Furthermore, hydrogel in a bead form was created using a droplet generating microfluidic device that has flow focusing microchannels. Reaction sensitivity was futher enhanced when biosensors were encapsulated in a bead form. In this case, phosphinothricin acetyltransferase (PAT), which is herbicide resistant protein that is related to marker of genetically modified (GM) crops was used as a model target. Using hydrogel bead biosensor presented in this study, PAT protein can be detected with naked eyes in a concentration as low as 20nM. This concept to detect low concentration of biomolecules can be potentially applied to other biosensors or analytical devices. 3239-Pos Board B616 Single Nucleotide Discrimination with Electro-Optical Nanopore Chan Cao, Yi-Tao Long. Ecust, Shanghai, China. Nanopore is an emerging technique, during the past two decades, has been explore to analysis the information about the identity, concentration, structure, and dynamics of the target molecules. However, the readout signal is mainly related on the tiny changes of ionic current as single molecules pass through the nanopore. Here, we develop the ultra-low current analyzer and combine plasmonic resonance scattering spectroscopy with nanopore technique, which could acquire the optical-electrical dual signals as a molecule pass though the nanopore. It provides more unique information of single biomolecule, which have a significant potential in life science. By virtue of this integrated strategy, we have realize single nucleotide discrimination, and even for the detecting the substituent difference in a molecule, such as DNA cytosine methylation. 1. Yi-Tao Long, Rui Gao, Chan Cao, Yi-Lun Ying, Da-Wei Li. CN203572764U[P]. 2. Gang Logan Liu, Yi-Tao Long, Yeonho Choi, Taewook Kang & Luke P Lee, Nat. Methods, 2007, 4(12), 1015-1017.
3240-Pos Board B617 Entropically Controlled Nanomechanical DNA Origami Devices Michael W. Hudoba1, Yi Luo2, Randy Patton1, Michael G. Poirier2, Carlos Castro1. 1 Mechanical & Aerospace Engineering, The Ohio State University, Columbus, OH, USA, 2Physics, The Ohio State University, Columbus, OH, USA. Structural DNA nanotechnology is a rapidly emerging field with immense potential for applications such as single molecule sensing, drug delivery, and manipulating molecular components. Major advances in the last decade have enabled the precise design and fabrication of DNA nanostructures with unprecedented geometric complexity; however, relative to natural biomolecular machines, the functional scope of DNA nanotechnology is limited by an inability to design dynamic mechanical behavior such as complex motion, conformational dynamics, or force generation. Inspired by approaches used in macroscopic machine design, we have recently developed methods to design structures with controllable 1D, 2D, or 3D motion. Moving beyond design of geometry or motion paths, we have recently developed structures with multiple thermally accessible states separated by well-defined energy barriers with tunable height. In particular, we have design a two-state device that transitions between compact and open states driven by thermal energy. We directly measured the dynamic behavior via single molecule fluorescence experiments, which show both the equilibrium distribution of states and the kinetics of switching between states can be tuned via design parameters that regulate the relative configurational space, or equivalently the entropy change, between states. Finally, we show that these dynamic structures can be used to probe local molecular scale forces, specifically depletion forces that result from entropic interactions with a crowding reagent. This work is the first demonstration of an ability to control the kinetics of DNA origami nanostructures down to the second scale and establishes a foundation for exploiting dynamic behavior of these devices to study physical interactions at the molecular scale.