Mahadhy Ally, Ståhl-Wernersson Eva, Mattiasson Bo, Hedström Martin
Department of Biotechnology, Lund University, P.O. Box 124, SE-22100 Lund, Sweden.
Biotechnol Rep (Amst). 2014 Jun 12;3:42-48. doi: 10.1016/j.btre.2014.06.002. eCollection 2014 Sep.
A capacitive DNA-sensor model system was used to monitor the capture of complementary single-stranded DNAs. The sensor chip consisted of a gold electrode, which was carefully insulated with a polytyramine layer and covalently tagged with 25-mer oligo-C. As low as 10 moles per liter of target oligo-G could be detected by injecting 250 μL of sample. Elevated temperature was used to reduce non-specific hybridization. Less than 10% of non-target 25-mer oligo-T interacted nonspecifically with the oligo-C probes when hybridization process was performed at 50 °C. Studying the relationship of length of the analyte to the signal strength, the output from the capacitive DNA-sensor increased to almost the double; from 50 to 88-nF cm, when a 25-mer oligo-G was used instead of a 15-mer. By sandwich hybridization at room temperature, it was possible to further increase the signal, from 78-nF cm for the target 50-mer oligo-G alone, to 114-nF cm.
一种电容式DNA传感器模型系统被用于监测互补单链DNA的捕获。传感器芯片由一个金电极组成,该电极用聚酪胺层仔细绝缘,并与25聚体寡聚-C共价标记。通过注入250μL样品,每升低至10摩尔的靶寡聚-G都能被检测到。使用升高的温度来减少非特异性杂交。当在50°C进行杂交过程时,不到10%的非靶25聚体寡聚-T与寡聚-C探针发生非特异性相互作用。研究分析物长度与信号强度的关系时,当使用25聚体寡聚-G代替15聚体时,电容式DNA传感器的输出增加到几乎两倍,从50 nF/cm增加到88 nF/cm。通过在室温下进行夹心杂交,有可能进一步增加信号,从单独的靶50聚体寡聚-G的78 nF/cm增加到114 nF/cm。