Tuesday November 14, 2006
4 PM
MIT- Building E25 (Whitaker College), Room 111

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"Lester Wolfe Workshop in Laser Biomedicine: Bio-Optics of DNA"
Prof. Amit Meller
Boston University

Analyzing DNA Dynamics and DNA Protein Interactions Using Nanopores and Single Molecule Fluorescence.

Negatively charged DNA molecules can be electrophoreticaly driven through a single nanometer-scale pore fabricated in thin silicon nitride films or through a membrane channel. When the nanopore diameter is made slightly above the size of double-stranded DNA (~2 nm) the DNA translocates through the pore in a single file, unfolded manner. Information about the translocation dynamics and DNA structure is obtained by probing the ionic current flowing through the nanopore during the passage of each biopolymer. This method is used in our lab for the analysis of nucleic acids' structure and DNA-protein interactions. Exploiting the native strong electric charge of nucleic acids, the nanopore sensor allows a high throughput analysis of unmodified and unconjugated individual biomolecules. We find that for unstructured polymers the DNA translocation is highly sensitive to its properties, such as its sequence and its direction of entry, and to physical parameters such as the driving field. Secondary structure in the polynucleotide leads to orders of magnitude slowing down in the translocation process. At large enough electrical field intensities structured DNA and RNA can be unzipped in a highly controlled way, and the unzipping kinetics is directly quantified. In this talk I will discuss our nanopore fabrication method, as well as a number of applications of the nanopore sensors for biomelcular analyses and our recent progress toward ultra fast single molecule DNA sequencing. Workshop Schedule

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