Thermal cycler and laminar flow workstations for gene discovery in non-model organisms

本提案中的设备需要用于支持基因发现研究，旨在阐明非模式生物中遗传性疾病的遗传基础，特别是那些具有区域保护、经济和管理重要性的基因发现研究。本研究的一个直接目标是利用全基因组连锁分析，结合生物信息学工具、实时PCR和微阵列表达分析的候选基因方法，发现与银狐遗传性增生性牙龈炎相关的基因和突变。狐狸的情况可以作为人类类似疾病的模型。此外，对这些类型疾病相关基因的研究有助于理解细胞生长和分化途径以及与功能相关的基因组变异的流行和分布。所需设备包括Eppendorf Mastercycler银块梯度热循环仪，用于扩增400个微卫星位点，进行全基因组连锁分析和候选基因的扩增和测序。此外，还需要两个Envirico桌面层流工作站，配有附件UV灯，用于无污染制备PCR主混合鸡尾酒和单独添加单个目标dna。额外的紫外线组件确保所有试剂，移液器和其他设备的无dna无菌。我目前正在建立一个实验室，致力于非模型、经济上重要的野生生物的基因发现研究和功能基因组学。所要求的设备项目是这一设施的基础。它们对实现我的研究目标至关重要，对任何基因组学导向的实验室都至关重要。他们还将在教育和培训学生方面发挥重要作用。

The equipment in this proposal is required to support gene discovery research that aims to elucidate the genetic basic of inherited diseases in non-model organisms, particularly those of conservation, economic and management importance regionally. One immediate goal of this research is to discover the gene(s) and mutation(s) associated with hereditary hyperplastic gingivitis in silver foxes using genome-wide linkage analysis in combination with a candidate gene approach that incorporates bioinformatics tools and Real Time PCR and microarray expression analyses. The fox condition serves as a model of a similar disease in humans. Furthermore, the investigation of genes involved in these types of diseases contribute to the understanding of cellular growth and differentiation pathways and the prevalence and distribution of functionally relevant genomic variation. The requested equipment includes an Eppendorf Mastercycler Silver Block Gradient Thermal Cycler required for the amplification of the 400 microsatellite loci necessary to conduct genome-wide linkage analysis and the amplification and sequencing of candidate genes. Additionally two Envirico Tabletop Laminar Flow Workstations with Accessory UV Lights are required for the contamination-free preparation of PCR master mix cocktails and the separate addition of individual target DNAs. The additional UV light components ensure the DNA-free sterility of all reagents, pipettors, and other equipment. I am currently in the process of establishing a laboratory dedicated to gene discovery research and functional genomics of non-model, economically-important wildlife organisms. The requested equipment items are the foundation for this facility. They are vital to the attainment of my research goals and essential to any genomics-oriented laboratory. They will also serve an important role in the education and training of students.