Evaluation of surface-enhanced resonance Raman scattering for quantitative DNA analysis
Evaluation of surface-enhanced resonance Raman scattering for quantitative DNA analysis
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DOI：
10.1021/ac035060c
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发表时间：
2004-01-15
影响因子：
7.4
通讯作者：
Graham, D
中科院分区：
文献类型：
--
作者：
Faulds, K;Smith, WE;Graham, D
使用染料标记生物物种已成为常见的做法，以帮助他们的检测，并在高稀释度的特定染料的立即阳性鉴定是一个关键的要求。在这里，通过表面增强共振拉曼散射（SERRS）的八个市售染料标记（BOX，罗丹明6 G，HEX，FAM，泰特，Cy 3，Cy 5，塔姆拉）连接到寡核苷酸链的检测报告。通过使用来自银纳米颗粒的SERRS产生独特的分子特异性光谱，可以容易地检测到八种标记中的每一种。优化条件以获得最佳信号增强，并获得低寡核苷酸浓度下的线性浓度图。在较高浓度下（高于10（-8）mol dm（-3）），除罗丹明6 G、泰特和FAM外，在浓度图中引入了曲率，在整个研究浓度范围内呈线性。得到的检测限低至0.5 fmol，如果分析较小的样品，则可能更低。还对含有表面活性剂吐温20的Tris-HCl缓冲液的作用进行了研究，以帮助防止超低浓度的寡核苷酸与样品容器的表面粘附。Tween 20允许获得所研究的每个标记的较低检测限。该研究表明，通常与寡核苷酸一起使用的不同染料可以在与荧光检测一起使用相同染料时不可能的浓度水平下给出定量SERRS。
The labeling of biological species using dyes has become common practice to aid in their detection, and immediate positive identification of specific dyes in high dilution is a key requirement. Here the detection by surface-enhanced resonance Raman scattering (SERRS) of eight commercially available dye labels (BOX, rhodamine 6G, HEX, FAM, TET, Cy3, Cy5, TAMRA) attached to oligonucleotide strands is reported. Each of the eight labels was easily detected by using the SERRS from silver nanoparticles to produce a unique, molecularly specific spectrum. The conditions were optimized to obtain the best signal enhancement, and linear concentration graphs at low oligonucleotide concentrations were obtained. At higher concentrations (above similar to10(-8) mol dm(-3)), curvature was introduced into the concentration graphs with the exception of rhodamine 6G, TET, and FAM, which gave linearity over the entire concentration range studied. Detection limits as low as 0.5 fmol were obtained, with lower possible if a smaller sample was analyzed. Investigation was also carried out into the effect of a Tris-HCl buffer containing the surfactant Tween 20 to aid in the prevention of surface adhesion of the oligonucleotides to the sample vessels at ultralow concentrations. The Tween 20 allowed lower detection limits to be obtained for each of the labels studied. This study shows that the different dyes commonly used with oligonucleotides can give quantitative SERRS at concentration levels not possible when the same dyes are used with fluorescence detection.