Construction of ultrahigh-density haplotyping devices using characteristics in nanospace

我们开发了由超高速基因组分析设备、超灵敏DNA检测设备和超小型反应室系统组成的超高密度单倍体基因组分析设备。在第一阶段，我们开发了这些组件器件中的基本技术。因此，优化了微通道内的纳米结构作为DNA筛选基质，实现了一种新的DNA标记方法的高灵敏度检测。此外，还证明了喷墨式注射方法确实可以作为DNA注射使用。第二年，我们研究了如何实现高密度微通道，因为直型微通道最适合制作高密度微通道阵列，优化了纳米结构和微通道的组合，并在直微通道上安装了喷墨式注射器。去年，对原型装置进行了最后的评估。结合已经开发的所有元器件，在芯片上制作了高集成度和高密度的器件作为原型器件。对于喷墨式喷油器，优化了施加电压、频率和波形，以减小单份油墨的体积分布。经过这一过程，我们可以将10Pl的DNA样品注入到直型微通道中，成功地进行了DNA的电泳分离。虽然高密度单倍型设备同样存在外围设备的物理干扰等问题，但我们证明了利用上述开发的技术可以实现高集成度和高密度的单倍型设备。

We have developed ultrahigh-density haplotyping devices composed of ultra-fast genome analysis device, ultra-sensitive DNA detection device, and ultra-small reaction chamber system for genome analysis. In the first yean essential techniques in these component devices were developed. As a result, nanostructures inside microchannel were optimized for DNA sieving matrix and high sensitive detection using a novel DNA labeling method was achieved. Furthermore, it was demonstrated that the inkjet-type injection method did work as a DNA injectr. In the seccond year we investigated how to army microchannel in high density Since straight type microchannel is most suitable for making high-density microchannel array, combination of nanostructures and microchannel was optimized and ink-jet-type injector was mounted on the straight miceothannel We elucidated that this combination worked well as a DNA analysis device. In the last year final evaluation of the prototype device was performed. Combining all components that have been developed so fat highly integrated and high-density device was fabricated on a chip as a prototype device. Regarding to inkjet-type injector applied voltage, frequency, and wave pattern was optimized to reduce the distribution of volume of a single aliquot. After this process, we could inject 10 pL DNA sample into the straight type microchannel and electrophoretic DNA separation was successfully done. Although high-density haplotyping devices has same problems such as physical interference of peripheral equipment, we demonstrated highly integrated and high-density haplotyping devices could be realized using the above developed techniques.