纳米金标记基因芯片技术的优化  

Optimization of Nanogold-labeled Microarray

作　　者：郑志伟[1] 常弘[1] 党华[2] 张国萍[1] 肖白玉[1] 杨登亮[1] 钟惟德[3] 

机构地区：[1]中山大学生命科学学院,广州510275 [2]中山大学附属第一医院黄埔院区,广州510700 [3]广州市第一人民医院,广州510180

出　　处：《生物技术通报》2010年第9期159-164,共6页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30872960);国家高技术研究发展计划项目("863"计划)(2006AA02A245;2007AA022001);广州市卫生局重点项目(2007-Zdi-05)

摘　　要：为了建立稳定的纳米金标记基因芯片技术,对点样液,预杂交,纳米金浓度,银染时间等进行了优化,并研究了该芯片方法的灵敏度。结果表明,使用50%DMSO作为点样液效果最好,预杂交会导致杂交信号降低70%,银染时间控制在15 min时候显色清晰且背景较低。该检测方法的灵敏度可达到80 fmol/L,可望在芯片检测领域得到更多的应用。To establish more stabilized nanogold-labeled microarray technology,we optimized the spotting buffer,the concentration of nanogold and the time of silver enhancement.Meanwhile,we studied the effect of pre-hybridization on hybridization signal,as well as the detection limit of the nanogold-labeled microarray.The results showed that the hybridization performed satisfactory specificity.Among the three kinds of spotting buffer,50%DMSO was the best.Pre-hybridization led to 70% loss of hybridization signal.The time of silver enhancement we chose was 15 minutes,at which the target signal was clear,while the background was low.The detection limit of nanogold-labeled microarray reach to 80 fmol/L,which was hopeful for variety of applications in the field of detection.

关 键 词：纳米金 生物素 链酶亲和素 寡核苷酸 银染显色 

分 类 号：Q789[生物学—分子生物学]