Gel and Blot Imager for Core Facility

申请资金用于分子动力学风暴840、Power Macintosh 7500、HP LaserJet 4M plus和6个成像屏幕，这些设备将放置在核心设施中，为生物、分子生物学与生物化学和化学三个部门服务。该仪器将由这些部门的六个主要用户、两个次要用户和其他临时用户的实验室使用。Storm 840成像放射性和荧光斑点和凝胶，具有五阶线性动态范围，放射性样品分辨率为250 M，荧光样品分辨率为500 M。该仪器将提供准确量化放射性和荧光斑点和凝胶的能力，这在卫斯理大学目前是不可能的。荧光能力将使荧光标签代替放射性标签在许多应用中使用。化学荧光技术替代化学发光技术将提供准确的样品定量和更长的样品寿命。风暴成像仪将用于定量蛋白质-DNA和蛋白质-RNA凝胶转移、测序凝胶、CAT测定、西部、北部和西南印迹、RNA足迹和足印、脉冲追踪标记实验、DNA复制、核苷酸结合、RNA酶保护和RT-PCR测定。这些方法将应用于几个不同的项目。这些研究包括：单个结构元件对RNA折叠的贡献；RNA-蛋白质相互作用的热力学和结构基础果蝇配对蛋白功能的解剖研究发育中中枢神经系统免疫球蛋白样分子的鉴定大肠杆菌蛋白定位机制的研究莱姆病诊断新抗原的鉴定酵母DNA复制与修复蛋白的生化分析细胞外基质和信号分子在畸胎瘤干细胞分化中的作用。卫斯理大学的配套资金将支付仪器费用的50%。

Funds are requested for a Molecular Dynamics Storm 840, Power Macintosh 7500, HP LaserJet 4M plus, and 6 imaging screens, to be placed in the core facility serving the three departments of Biology, Molecular Biology & Biochemistry, and Chemistry. The instrument will be used by the laboratories of six major users, two minor users, and other occasional users in these departments. The Storm 840 images radioactive and fluorescent blots and gels with five orders of linear dynamic range, and a resolution of 250 M for radioactive samples, and 500 M for fluorescent samples. This instrument will provide the ability to accurately quantify radioactive and fluorescent blots and gels, which is not currently possible at Wesleyan University. The fluorescent capability will enable the use of fluorescent labels instead of radioactive labels in many applications. Substitution of chemifluorescent techniques for chemiluminescent techniques will provide both accurate quantitation of samples and longer sample lifetime. The Storm imager will be used to quantify protein-DNA and protein-RNA gel shifts, sequencing gels, CAT assays, Western, Northern, and Southwestern blots, RNA footprints and toeprints, pulse-chase labeling experiments, processive DNA replication, nucleotide binding, RNase protection, and RT-PCR assays. These methods will be applied to several different projects. These include studies of: Contribution of individual structural elements to the folding of RNA; Thermodynamic and structural basis of RNA- protein interactions; Dissection of Drosophila Paired proteil function; Identification of imm unoglobulin-like molecules in he developing CNS; Mechanisms of protein localization in scherichia coli; Identification of novel antigens for Lyme disease diagnosis; Biochemical analysis of yeast DNA replication and repair proteins; and The role of extracellular matrix and signaling molecules in teratocarcinoma stem cell differentiation. Matching funds from Wesleyan University will pay for 50% of the cost of the instrument.