October 21, 2000Analytical Chemistry
A Fluorescence-Based Method for Determining the Surface Coverage and Hybridization Efficiency of Thiol-Capped Oligonucleotides Bound to Gold Thin Films and Nanoparticles
Authors
Discussion
Loading...
Member takes
Implication
In vitro study reveals higher DNA coverage but lower hybridization on gold nanoparticles than planar films, indicating strategies to optimize nanoparticle-based DNA biosensors.
Key Points
- To measure and compare the surface coverage and hybridization efficiency of thiol-modified oligonucleotides bound to gold nanoparticles versus planar gold thin films using a fluorescence-based method.
- Quantified thiol-derivatized single-stranded oligonucleotides on gold nanoparticles and planar gold thin films using fluorescence assays.
- Evaluated the impact of progressive electrolyte addition during deposition, oligonucleotide spacer sequences, and coadsorbed diluent strands on surface packing and hybridization.
- Gold nanoparticles demonstrated higher oligonucleotide surface coverage than planar gold thin films (34 ± 1 pmol/cm² vs 18 ± 3 pmol/cm²), but lower initial hybridization efficiency (1.3 ± 0.3 pmol/cm² [4%] vs 6 ± 2 pmol/cm² [33%]).
- Incorporating spacer sequences increased hybridization efficiency on nanoparticles from approximately 4% to 44%, while diluent oligonucleotides allowed precise tuning of target-bound strand density.
Cite This Study
Demers et al. (2000) studied this question.