Williams J.G., Reynolds B.L., Keefe K., Anderson J.P.
LI-COR Biosciences, US
Keywords: fluorescent nanoparticles, nucleotides, polymerase, structural DNA, SYBR
Structural DNA nanoparticles functionalized with phosphate-linked nucleotide triphosphates (dNTPs) were tested as a source of dNTPs for DNA polymerase. The particles were prepared by strand-displacement polymerization from a self-complementary circular template. An intercalating dye (SYBR-101) was joined to the gamma-phosphate of a dNTP by a PEG linker, and the dye-dNTP conjugate was covalently bonded to the nanoparticles. Imaged by AFM, these functionalized particles appear as condensed fuzzy balls with diameters between 50-150 nm. They emit a bright fluorescent signal, detected in 5 msec exposures with a signal-to-noise of 50 when imaged using a TIR fluorescence microscope. A DNA polymerase, adapted by directed evolution to the efficient utilization of phosphate-modified dNTPs, inefficiently utilizes dNTPs supplied from the nanoparticles.
Journal: TechConnect Briefs
Volume: 2, Technical Proceedings of the 2007 NSTI Nanotechnology Conference and Trade Show, Volume 2
Published: May 20, 2007
Pages: 239 - 242
Industry sectors: Advanced Materials & Manufacturing | Medical & Biotech
Topics: Biomaterials, Materials Characterization & Imaging
ISBN: 1-4200-6183-6