Affinity capillary electrophoresis with a DNA-nanoparticle conjugate as a new tool for genotyping.
We have developed a novel method for genotyping based on free solution affinity capillary electrophoresis. We prepared DNA-nanoparticle conjugates by mixing biotin-modified DNA and NeutrAvidin-modified polystyrene nanoparticles; this mixture was then injected into a capillary. Subsequently, we injected the fluorescent-labeled sample DNAs into the capillary, applied the voltage, increased its temperature after 7 min, and detected the fluorescence at its anodic end. This novel method was applied for genotyping human c-K-ras, and the three genotypes were definitely distinguishable with high reproducibility. This method can be easily automated, and it is useful for high-throughput gene mutation analysis.
Adachi
Ken
K
Department of Chemical Engineering, Waseda University, 3-4-1 Ohkubo, Shinjuku-ku, Tokyo 169-8555, Japan.
Noda
Naohiro
N
Nakashige
Makoto
M
Tsuneda
Satoshi
S
Kanagawa
Takahiro
T
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D017931:DNA Primers; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes; D011905:Genes, ras; D005838:Genotype; D036103:Nanotechnology
D017931:DNA Primers; D005456:Fluorescent Dyes; D004247:DNA