...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Electroanalysis >Scanning electrochemical microscopy (SECM) based detection of oligonucleotide hybridization and simultaneous determination of the surface concentration of immobilized oligonucleotides on gold
【24h】

Scanning electrochemical microscopy (SECM) based detection of oligonucleotide hybridization and simultaneous determination of the surface concentration of immobilized oligonucleotides on gold

机译：基于扫描电化学显微镜（SECM）的寡核苷酸杂交检测以及同时测定金上固定寡核苷酸的表面浓度

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The direct mode of scanning electrochemical microscopy (SECM) was used for the local deposition of oligonucleotide (ODN) patterns on thin gold films and the generation-collection (GC) mode was applied for the determining the amount of surface-accessible oligonucleotides. The local deposition was achieved through the micrometer-sized formation of a conducting polymer bearing 15(mcr) single-stranded oligonucleotide strands. After the interaction of the oligonucleotide with its biotin-labeled complimentary strand, streptavidin was bound. The molecular assembly was completed by linking biotin-labeled P-galactosidase from Escherichia coli to the streptavidin. The activity of the linked beta-galactosidase was mapped with SECM in the GC mode by monitoring the oxidation of p-aminophenol (PAP) formed in the enzyme-catalyzed hydrolysis of p-aminophenyl-beta-D-galactopyranoside. The feedback effect due to recycling of the reaction product at the gold surface was analyzed. It was shown experimentally that this effect becomes insignificant at ultramicroelectrode (UME)-substrate distances larger than 3 UME radii. The flux of formed PAP allowed the determination the surface density of accessible oligonucleotide strands in the functionalized polymer. It was shown that that thicker pyrrole/ODN-Pyrrole polymer films do not lead to a significantly increased accessible ODN surface concentration.
机译：扫描电化学显微镜（SECM）的直接模式用于在金薄膜上局部沉积寡核苷酸（ODN）模式，而世代收集（GC）模式用于确定表面可及的寡核苷酸的数量。通过微米尺寸的带有15（mcr）单链寡核苷酸链的导电聚合物的形成来实现局部沉积。寡核苷酸与其生物素标记的互补链相互作用后，链霉亲和素被结合。通过将大肠杆菌的生物素标记的P-半乳糖苷酶与链霉亲和素连接起来，完成了分子组装。通过监测在对氨基苯酚-β-D-吡喃半乳糖苷的酶催化水解过程中形成的对氨基苯酚（PAP）的氧化，在GC模式下用SECM绘制了连接的β-半乳糖苷酶的活性图。分析了由于反应产物在金表面的再循环而产生的反馈效应。实验表明，在大于3 UME半径的超微电极（UME）-基底距离处，这种作用变得微不足道。形成的PAP的通量允许确定官能化聚合物中可及的寡核苷酸链的表面密度。结果表明，较厚的吡咯/ ODN-吡咯聚合物薄膜不会导致可及的ODN表面浓度显着增加。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号