一种基于SU8聚合物的基因测序芯片

基因测序技术极大地推动了生物学和医学研究的发展. 结合了焦磷酸测序 原理及阵列式微反应池芯片的高通量测序仪在从头测序和宏基因组测序方面有着不可替代的作用. 本文首次提出并研制了一种基于SU8聚合物的基因测序芯片. 选择了高传输效率、 低耦合损耗的光纤面板作为基片, 通过改善SU8均匀性及释放应力, 在光纤面板上成功制备出百万数量级阵列式微反应池; 设计并制作侧壁镀膜装置, 实现了SU8阵列式微反应池侧壁选择性光学薄膜蒸镀, 有效地提高了微反应池的光学隔离度, 将相邻微反应池之间的光串扰率平均值从25%降低到了1%, 满足了高通量焦磷酸测序对测序芯片独立并行传输弱光信号的要求. 基于SU8聚合物的基因测序芯片制备工艺简单、成本低廉, 具有良好的应用前景. 关键词： DNA测序 SU8 微反应池 光纤面板

Fabrication of SU8-based chip suitable for genomic sequencing

DNA sequencing technology has markedly advanced the development of biological and medicinal sciences. High-throughput pyrosequencing instruments that combine the pyrosequencing with microfabricated high-density picoliter reactors have been proved to be suitable for de novo sequencing and metagenome sequencing. In the present work, we report on an alternative sequencing chip consisting of hundreds of thousands of picoliter sized honeycombed SU8 reaction vessels on a fiber-optic slide by lithography technique for high-throughput pyrosequencing instruments. Highly reproducible fabrication process of SU8 sequencing chip is achieved through the improvement on SU8 film thickness uniformity and relaxation of SU8 residual stress during fabrication. To achieve the optical isolation required for SU8 reaction wells, metal film is selectively deposited on the side walls of the reaction vessels by reformating vacuum coating. With the metal coating, the average value of optical cross talking between SU8 reaction vessels is reduced from 25% to 1%. The SU8 sequencing chip demonstrates an excellent light transmission characteristic and meets the need of pyrosequencing application.
Keywords： DNA sequencing SU8 picoliter well fiber optic plates