Integrated Robotics for Microarray Assembly (IRMA)

该应用程序需要用于定制cDNA和基因组文库的基因组规模组装的仪器，用于微阵列转录物分析和模式生物的功能和遗传筛选。该仪器提供了一个集成的克隆组装系统，称为微阵列组装集成机器人（IRMA），包括：1)用于回收和组装大型细菌和酵母克隆文库的机器人菌落拾取器；2) 96孔培养箱，用于小体积、高密度培养大型克隆；3)用于大规模DNA纯化和PCR扩增反应组装的液体处理机器人；4)用于PCR扩增的高容量热循环仪，用于序列分析和微阵列目标的生产。IRMA仪器将被安置在宾夕法尼亚大学的微阵列核心设施中，在核心实验室人员的监督下，为克隆管理和微阵列应用提供一个高效和生产的技术专业知识和核心设施中可用的下游仪器。参与研究的8位研究人员有资金充足的研究项目，研究复杂发育过程的遗传调控，包括干细胞规范、身体计划模式和轴突引导，使用多种模式生物，包括小鼠、斑马鱼、小鸡、果蝇和拟南芥。这种IRMA仪器将是这些项目的重要组成部分，使研究人员能够组装大量的克隆，以追求基因组规模的功能和基于表达的筛选，这些筛选需要定义介导这些发育过程的基因网络。

This application requests instrumentation for genome scale assembly of custom cDNA and genomic libraries for microarray transcript profiling analysis and for functional and genetic screens for model organisms. This instrumentation provides an integrated clone assembly system, referred to as Integrated Robotics for Microarray Assembly (IRMA), including: 1) a Robotic Colony Picker for recovering and assembling large libraries of bacterial and yeast clones; 2) an Incubating Growth Chamber for small volume, high density growth of large clone collections in a 96-well format; 3) a Liquid Handling Robot for large scale DNA purification and assembly of PCR amplification reactions; and 4) a High Capacity Thermocycler for PCR amplifications for sequence analysis and microarray target production. The IRMA instrumentation will be housed in the University of Pennsylvania Microarray Core Facility, under the supervision of Core lab personnel, providing an efficient and productive interface with the technical expertise and downstream instrumentation available in the Core Facility for clone management and microarray applications. The eight participating investigators have well-funded research programs investigating genetic regulation of complex developmental processes, including stem cell specification, body plan patterning, and axon guidance, using a variety of model organisms, including Mouse, Zebra fish, Chick, Drosophila, and Arabidopsis. This IRMA instrumentation will be an essential component of these projects, allowing investigators to assemble the large set of clones to pursue genome-scale functional and expression-based screens that are required to define the gene networks that mediate these developmental processes.