Core--Genotyping

该组件包含提案的基因分型和DNA收集部分。我们建议在人类遗传学中心目前使用的现有寡核苷酸连接试验的基础上，开发一种高通量的SNP基因分型试验。奥拉测定具有高度灵活、成本有效、准确、适合小型大型实验室的优点，因为不需要昂贵的设备，并且可以多路复用。我们将研究使用多重PCR进行奥拉分析的动力学，开发常规奥拉多重PCR的标准方案，并使该程序适用于8通道和96通道毛细管测序仪。这将是基于我们非常成功的Floughenes等位基因静态扫描技术（FASST），我们在过去的一年中已经使用该技术对700，000个微卫星进行了基因分型。 一旦开发出该程序，我们将使用它来测试组件3，SNP发现中开发的SNP，来自表达研究中鉴定的基因。此外，我们将研究是否从表达供体获得的DNA上的基因分型SNP可以有效地减少在更大的心脏数据集上进行基因分型的候选基因的数量。将收集2000个个体的数据集，以观察这些表达/关联方法对复杂疾病是否会成功。

This component contains the genotyping and DNA collection portions of the proposal. We propose building upon the existing oligonucleotide ligation assay currently in use in the Center for Human Genetics to develop a high-throughput SNP genotyping assay. The OLA assay has the advantage of being highly flexible, cost effective, accurate, good for small large labs, as no expensive equipment is necessary, and can be multiplexed. We will investigate the dynamics of using PCR multiplex for OLA analysis, develop standard protocol for routine OLA multiplexes, and adapt the procedure to both 8 and 96 channel capillary sequencers. This will be based on our highly successful Flourescent Allele Static Scanning Technique (FASST), which we have used to genotype 700,000 microsatellite in the past year. Once the procedure is developed, we will use it to test the SNPs developed in Component 3, SNP Discovery, from the genes identified in the expression studies. In addition, we will investigate whether genotyping SNPs on DNA obtained from the expression donors can efficiently reduce the number of candidate genes genotyped on a larger cardiac data set. This data set of 2000 individuals will be collected to observe if these expression/association approach to complex disorders will be successful.