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TITLE:
NanoSelectionTM -- a Single-Cycle Method, Based on a Combined
AFM/FM Instrument, to Select Individual Aptamer Species from a Pool of
Random Oligonucleotides
SPEAKER:
Lu Peng,
TIME: Thursday Nov. 30, 2006
at 3:30*
PLACE: Olin 101
Wake Forest University
*Note unusual time.
A new method to select individual aptamer species from a pool of random molecules based on the aptamer's affinity to a target molecule---- "NanoSelectionTM technology" is introduced. At the core of this method is instrumentation that combines an atomic force microscope and an inverted optical fluorescence microscope. In this method, target molecules are attached to a cover slip. A library of fluorescent oligos attached to a small bead (for AFM detection and extraction) is then dispensed to the target area. High-affinity, target-specific aptamers will bind tightly to the target for prolonged periods resulting in a strong fluorescence signal (and potentially fluorescence resonance energy transfer (FRET) with accept-labeled oligos and donor-labeled targets) and making them resistant to washes. The AFM is then directed to the coordinates of the fluorescence signal, and a high resolution image of the area is obtained. Subsequently, the AFM tip is used to extract the bead plus the attached oligo. The extracted oligo is amplified by small-copy-number PCR, followed by a biochemical analysis.