Adachi Ken, Noda Naohiro, Nakashige Makoto, Tsuneda Satoshi, Kanagawa Takahiro
Department of Chemical Engineering, Waseda University, 3-4-1 Ohkubo, Shinjuku-ku, Tokyo 169-8555, Japan.
J Chromatogr A. 2006 Mar 24;1109(2):127-31. doi: 10.1016/j.chroma.2005.12.110. Epub 2006 Jan 30.
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.
我们开发了一种基于自由溶液亲和毛细管电泳的新型基因分型方法。我们通过将生物素修饰的DNA与中性抗生物素蛋白修饰的聚苯乙烯纳米颗粒混合来制备DNA-纳米颗粒缀合物;然后将该混合物注入毛细管中。随后,我们将荧光标记的样品DNA注入毛细管,施加电压,7分钟后升高温度,并在其阳极末端检测荧光。这种新方法被应用于人类c-K-ras基因分型,三种基因型能够被明确区分,且具有高重现性。该方法易于自动化,可用于高通量基因突变分析。