交联型丙烯酸酯聚合物分子层用于固定寡核苷酸分子初探

The study of oligonucleotides immobilization film prepared with acrylic acid-co-acrylamide copolymer

目的探讨在显微玻片上衍生聚合物分子层用于固定寡核苷酸分子的方法,为基因芯片制备和应用提供技术支持。方法利用交联聚合反应,在显微玻片表面衍生丙烯酸酯共聚物分子层,结合基因芯片技术制备寡核苷酸芯片。利用荧光标记的样品和CMT芯片杂交系统对所制备的芯片进行杂交实验,检测寡核苷酸在芯片表面的固定性能,并进行对照实验。结果杂交后的荧光检测表明寡核苷酸在丙烯酸酯共聚物表面上杂交点阵的荧光信号多,信号强度强,杂交点圆润、同一性好。结论利用化学交联法制备的芯片表面能用于寡核苷酸的固定且处理过程简单,为进一步运用到生物芯片制作打下了良好的基础。

Objective To explore the key technologies in DNA microarray fabrication is immobilizing DNA or oligonucleotide efficiently on modified surface. Methods The glass slides were modified with acrylic acid-co-acrylamide copolymer and EDC/NHS by covalent linkage method. Results A covalent immobilization of unmodified oligonucleotide on the glass surface was achieved. The platform was prepared for oligo-microarray fabrication. Hybridization experiments were showed that the modified surfaces have high fluorescence intensity, low background in contrast unmodified slide. The uniform, steady and regular shaped signal spots of the microarrays indicated that the prepared surfaces were satisfactory in immobilizing unmodified oligonucleotides. Conclusion The process of the surface preparation is simple and cost effective and can be used for attachment of protein, PNA et cetra.