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DNA Oligonucleotide Synthesis in Mesoporous Silicon for Biosensing Applications

机译:DNA寡核苷酸在介孔硅中合成生物传感应用

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We report a method for improving the sensitivity of label-free optical biosensors based on in-situ synthesis of DNA probes within porous silicon structures. The stepwise attachment of up to 15mer probes inside 30 nm mesopores was accomplished through a series of phosphoramidite reactions. In this work, a porous silicon waveguide was utilized as the sensor structure. Synthesis of DNA probe, as well as sensing of target DNA, was verified by monitoring the change in effective refractive index of the porous silicon waveguide through angle-resolved attenuated total reflectance measurements. The average resonance shift per oligo of 0.091 ° during stepwise synthesis corresponds to surface coverage slightly less than 50%, according to theoretical models. When compared with the traditional method of direct attachment of pre-synthesized oligonucleotide probes, the sequential phosphoramidite method resulted in an approximately four-fold increase in DNA probe attachment. This increased surface coverage by DNA probes increases the likelihood of target molecule binding, leading to improved sensitivity for bio-molecule detection. Exposure to a 50μM solution of target 8-base DNA in deionized water produced a 0.4236° change in the waveguide resonance angle. Nanomolar detection limits for small molecule sensing are realizable with this sensor scheme.
机译:我们报告用于改进基于原位多孔硅结构内合成DNA探针的无标记的光学生物传感器的灵敏性的方法。达15聚体探针内部30nm的中孔的阶梯状附着是通过一系列的亚磷酰胺的反应来完成。在这项工作中,多孔硅波导被用作传感器的结构。 DNA探针,以及靶DNA的检测的合成,通过监测通过角分辨衰减全反射测量在多孔硅波导的有效折射率的变化验证。期间逐步合成对应于大于50%稍少的表面覆盖每0.091°寡平均共振位移,按照理论模型。当与预先合成的寡核苷酸探针直接连接的传统方法相比,顺序亚磷酰胺法导致DNA探针附件的大约增加四倍。通过DNA探针该增加的表面覆盖度增大目标分子的结合的可能性,从而导致用于生物分子检测改进的灵敏度。暴露于在去离子水中靶8-碱基的DNA的50μM溶液制备在波导共振角的0.4236°的变化。对于小分子传感纳摩尔检测限是可实现的与该传感器方案。

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