Quantitative evaluation of oligonucleotide surface concentrations using polymerization-based amplification.
Quantitative evaluation of oligonucleotide surface concentrations using polymerization-based amplification.
复制标题
使用基于聚合的扩增对寡核苷酸表面浓度进行定量评估。
DOI：
10.1007/s00216-008-2259-6
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发表时间：
2008-09
影响因子：
4.3
通讯作者：
Bowman, Christopher N.
中科院分区：
文献类型：
--
作者：
Hansen, Ryan R.;Avens, Heather J.;Shenoy, Raveesh;Bowman, Christopher N.
关键词：
在分子诊断技术中发现的无数应用中，最小多核苷酸浓度的定量评估已经成为关键分析。因此，开发灵敏、定量且可用于即时检测的高通量、非酶测定有利于常规实施。在这里，我们开发了一种非酶的方法，通过耦合调节量的聚合物生长互补的生物分子结合在基于阵列的生物芯片上的标记的DNA目标的表面浓度进行定量。在20-220 nm范围内的聚合物膜厚度测量值随标记的DNA表面浓度的变化而变化，超过两个数量级，定量下限为60个分子/μm2（约106个靶分子）。在努力开发这种扩增方法与基于荧光的表征方法的兼容性中，还评估了将荧光纳米颗粒掺入聚合物膜中。荧光信号的所得增益使得能够使用适合于护理点设置的检测仪器进行定量。基于聚合的扩增用于3'生物素化寡核苷酸表面浓度的定量评价
Quantitative evaluation of minimal polynucleotide concentrations has become a critical analysis among a myriad of applications found in molecular diagnostic technology. Development of high-throughput, nonenzymatic assays that are sensitive, quantitative and yet feasible for point-of-care testing are thus beneficial for routine implementation. Here, we develop a nonenzymatic method for quantifying surface concentrations of labeled DNA targets by coupling regulated amounts of polymer growth to complementary biomolecular binding on array-based biochips. Polymer film thickness measurements in the 20–220 nm range vary logarithmically with labeled DNA surface concentrations over two orders of magnitude with a lower limit of quantitation at 60 molecules/μm2 (∼106 target molecules). In an effort to develop this amplification method towards compatibility with fluorescence-based methods of characterization, incorporation of fluorescent nanoparticles into the polymer films is also evaluated. The resulting gains in fluorescent signal enable quantification using detection instrumentation amenable to point-of-care settings. Polymerization-based amplification for quantitative evaluation of 3’ biotinylated oligonucleotide surface concentrations
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影响因子：
14.9
作者：
Peterson, AW;Heaton, RJ;Georgiadis, RM
通讯作者：
Georgiadis, RM
影响因子：
14
作者：
Kizilel, S;Sawardecker, E;Pérez-Luna, VH
通讯作者：
Pérez-Luna, VH
影响因子：
6.2
作者：
Hansen, Ryan R.;Sikes, Hadley D.;Bowman, Christopher N.
通讯作者：
Bowman, Christopher N.
影响因子：
41.2
作者：
Sikes, Hadley D.;Hansen, Ryan R.;Bowman, Christopher N.
通讯作者：
Bowman, Christopher N.
影响因子：
7
作者：
Pastinen, T;Raitio, M;Syvänen, AC
通讯作者：
Syvänen, AC