A SNP-BASED, HIGH-THROUGHPUT, LOW-COST GENOTYPING METHOD

该提案的目的是开发和优化设计用于产生使用单核苷酸多态性（SNP）作为遗传标记的新型、低成本、高通量基因分型方法的方法。该方法依赖于创新性地使用降低复杂性的基因组、微阵列技术和等位基因特异性寡核苷酸（阿索）杂交技术，以允许快速有效地鉴定SNP并同时对数千个个体进行基因分型。该方法允许从单个PCR反应中分析超过100个SNP。这种基于SNP的基因分型方法的发展最终将允许对糖尿病和哮喘等复杂性状进行遗传解剖。初步实验已经产生了复杂性降低的基因组，其允许有效鉴定超过50个SNP，这些SNP在来自多个CEPH家族的DNA上的阿索杂交中具有50%的成功率。 该提议将允许鉴定在阿索杂交中具有大于80%成功率的最大数目的SNP。此外，该提议旨在鉴定大约100个独特的SNP并确定其基因组图谱位置和等位基因频率。拟议的商业应用：高通量、低成本的基因分型技术将支持大规模人群研究，以从基因上剖析复杂的性状，如糖尿病、肥胖和心脏病。这对于理解和诊断许多常见疾病以及发现诺埃尔治疗方法具有重要的商业应用。这项技术是理想的药物基因组学研究，以确定候选人群可能受益于诺埃尔治疗药物。此外，它提供了一种快速，低成本的方法，数字DNA指纹的军事和刑事鉴定和法医应用。

The objective of this proposal is the development and optimization of methodologies which are designed to generate a novel, low-cost, high- throughput method of genotyping using single nucleotide polymorphisms (SNPs) as genetic markers. This method relies on the innovative use of reduced complexity genomes, microarray technology and allele-specific oligonucleotide (ASO) hybridization techniques to allow the rapid and efficient identification of SNPs and the simultaneous genotyping of thousands of individuals. This method allows the analysis of over 100 SNPs from a single PCR reaction. The development of such a SNP-based genotyping method will eventually allow the genetic dissection of complex traits such as diabetes and asthma. Preliminary experiments have generated a reduced complexity genome that has allowed the efficient identification of over 50 SNPs which have a 50% success rate in ASO hybridization on DNA derived from multiple CEPH families. This proposal will allow the identification of a maximum number of SNPs having a greater than 80% success rate in ASO hybridizations. Additionally this proposal is designed to identify approximately 100 unique SNPs and determine their genomic map location and allele frequency. PROPOSED COMMERCIAL APPLICATIONS: A high throughput, low-cost genotyping technology will support large population studies to genetically dissect complex traits such as diabetes, obesity and heart disease. This has significant commercial applications for understanding and diagnosis of many common diseases as well as the discovery of noel therapeutic treatments. This technology is ideal for pharmacogenomic studies to identify candidate populations likely to benefit from noel therapeutic agents. Additionally it offers a rapid, low- cost approach for digital DNA fingerprinting for military and criminal identification and forensic applications.