Liu Guodong, Lee Thomas M H, Wang Joseph
Department of Chemical Engineering, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-6006, USA.
J Am Chem Soc. 2005 Jan 12;127(1):38-9. doi: 10.1021/ja043780a.
A bioelectronic method for coding unknown single nucleotide polymorphisms (SNPs) based on the use of different encoding nanocrystals is described. Four such nanocrystals, ZnS, CdS, PbS, and CuS, linked to the adenosine, cytidine, guanosine and thymidine mononucleotides, respectively, are sequentially introduced to the DNA hybrid-coated magnetic-bead solution. Each mutation captures via base pairing different nanocrystal-mononucleotide conjugates, and yields a characteristic multipotential voltammogram, whose peak potentials reflect the identity of the mismatch. The mismatch recognition events are being amplified by the metal accumulation feature of the stripping voltammetric transduction mode. Each of the eight possible one-base mismatches can thus be identified in a single voltammetric run. The use of nanocrystal tracers for detecting two known mutations in a single DNA target is also illustrated in connection to nanocrystals linked to two nucleotides along with a single voltammetric run. The protocol presented should facilitate the rapid, simple, low-cost, and high throughput screening for SNPs.
描述了一种基于使用不同编码纳米晶体对未知单核苷酸多态性(SNP)进行编码的生物电子方法。分别与腺苷、胞苷、鸟苷和胸苷单核苷酸相连的四种这样的纳米晶体,即硫化锌、硫化镉、硫化铅和硫化铜,被依次引入到DNA杂交包被的磁珠溶液中。每个突变通过碱基配对捕获不同的纳米晶体 - 单核苷酸共轭物,并产生特征性的多电位伏安图,其峰值电位反映错配的身份。错配识别事件通过溶出伏安传感模式的金属积累特性进行放大。因此,在一次伏安测量中可以识别出八种可能的单碱基错配中的每一种。还结合与两个核苷酸相连的纳米晶体以及一次伏安测量,说明了使用纳米晶体示踪剂在单个DNA靶标中检测两个已知突变的情况。所提出的方案应有助于对SNP进行快速、简单、低成本和高通量筛选。