HUMAN DIALLELIC INSERTION/DELETION OF POLYMORPHISMS

描述：（申请人摘要）人类DNA多态性发现 在许多研究和临床试验领域的应用，特别是 疾病相关基因的连锁图谱 短串联重复 多态性目前是这些应用的选择标记， 但双等位基因多态性可能最终取代STRP， 小型化DNA阵列（DNA芯片）更有效的分析。 目标 该研究项目是第一个，开发1,500个新的人类双等位基因 短插入/缺失类的多态性，第二，调查 新办法，并加强现有办法， 插入/缺失多态性对流通DNA基因传感器的影响，第三， 逐步引入基于芯片的双等位基因分析， 基因多态性的基因分型工厂。 双等位基因短 插入/缺失多态性具有许多优于碱基 取代多态性，尤其是改良杂交的取代多态性 等位基因之间的区别。 新的推定插入/缺失 多态性将通过比较公开可获得的重叠 基因组和编码DNA序列。 将确认推定的多态性 实验室类型。 将在以下组中测量等位基因频率： 具有美洲、欧洲、非洲和亚洲血统的个体。 Drs. 橡树岭国家实验室的贝蒂和多克蒂茨开创了 新型多孔DNA芯片材料，特别是毛细管玻璃。 的 多孔芯片支持增加的杂交反应动力学相比， 扁平薯片 协议将得到加强，以生产芯片， 将DNA靶与芯片杂交，并用于检测 杂交DNA 许多新颖的多态性分析方法， 杂交靶PCR扩增的要求将是 探讨了 用于芯片排列和阅读的仪器将得到改进。 最后，用于分型短插入/缺失多态性的DNA芯片将在 逐渐引入马什菲尔德的基因分型工厂， 疾病基因连锁分析中的STRP分型

DESCRIPTION: (Applicant's abstract) Human DNA polymorphisms find application in many areas of research and clinical testing, particularly in the linkage mapping of genes responsible for disease. Short tandem repeat polymorphisms are currently the markers of choice for these applications, but diallelic polymorphisms will likely eventually replace STRPs because of more efficient analysis on miniaturized DNA arrays (DNA chips). Goals of this research project are first, to develop 1,500 new human diallelic polymorphisms of the short insertion/deletion class, second, to investigate new approaches and to enhance existing approaches for the analysis of the insertion/deletion polymorphisms on flow through DNA genosensors, and third, to gradually introduce the chip-based analysis of the diallelic polymorphisms into the genotyping factory at Marshfield. Diallelic short insertion/deletion polymorphisms have a number of advantages over base substitution polymorphisms, especially that of improved hybridization discrimination between alleles. New, putative insertion/deletion polymorphisms will be identified by comparing publicly available overlapping genomic and coding DNA sequences. Putative polymorphisms will be confirmed by laboratory typing. Allele frequencies will be measured in groups of individuals with American, European, African, and Asian ancestry. Drs. Beattie and Doktycz at Oak Ridge National Laboratory have pioneered work on novel, porous DNA chip materials, particularly capillary channel glass. The porous chips support increased hybridization reaction kinetics compared to flat chips. Protocols will be enhanced for production of chips, for hybridization of DNA targets to the chips, and for detection of the hybridized DNA. A number of novel polymorphism analysis methods that bypass the requirement for PCR amplification of hybridization targets will be explored. Instrumentation for chip arraying and reading will be improved. Finally, DNA chips for typing short insertion/deletion polymorphisms will be gradually introduced into the genotyping factory at Marshfield to supplement STRP typing in linkage analysis of disease genes.