Picking and Gridding Q Bot System

描述（由申请人提供）：本提案的目的是寻求资金购买机器人高分辨率拾取和网格化Q-bot系统。大文库如cDNA文库或噬菌体展示文库的操作是相当麻烦的，并且充满了克隆损失或污染点。使用机器人拾取器和格栅可以缓解这些问题。目前，在大克利夫兰/东北俄亥俄州地区还没有这样的文书。这种机器人挑选和网格化仪器将大大提高克利夫兰临床基金会（CCF）勒纳研究所内基因表达实验室和临床医学核心DNA实验室（DLCM）的cDNA阵列能力。所要求的仪器也将用于促进和推进该地区内的机构（CCF和凯斯西储大学）的研究计划。主要用户都有许多NIH资助的研究赠款，这些基金具有可以最好地使用该仪器系统的菌落殖民地/噬菌体挑选和重新排列/樱桃挑选或高密度排列特征进行的组成项目。所有主要用户的项目都需要大量的需求来操纵大型噬菌体或cDNA文库。主要用户的项目范围很广，包括：1）鉴定病变组织中差异表达的肿瘤抑制基因或癌基因; 2）鉴定复杂生物蛋白质混合物中具有确定蛋白质组分的相互作用蛋白质; 3）蛋白质：蛋白质相互作用结构域的表征;以及4）识别响应于生物恐怖生物制剂的改变的细胞信号传导途径和基因表达模式。Q-bot系统提供了满足这些不同需求的多功能性、分辨率和功能。此外，所要求的仪器系统将用于满足偶尔使用者的需要。这项服务将在我们现有的分子生物技术核心内进行，该核心通过进行蛋白质测序、肽合成和相关服务为研究人员提供支持。由于人类基因组计划，DNA排列设备的使用量大大增加。使用Q-bot，CCF的DNA阵列设施将能够生成高度多样化的阵列类型，并维持和复制其经验证的40 K人类和15 K小鼠文库的宝贵cDNA序列。CCF已投入大量资源发展临床医学DNA实验室和分子生物技术核心，包括监督其运作的人员，以及确保其长期运作和维护的财政计划。总之，获得所要求的高分辨率拾取和网格化Q-bot系统将对该地区多个机构（包括CCF和CWRU）的生物医学研究项目的进展和未来发展产生重大影响。

DESCRIPTION (provided by applicant): The purpose of this proposal is to seek funds to purchase a robotic high-resolution picking and gridding Q-bot system. Manipulation of large libraries such as cDNA libraries or phage display libraries is quite cumbersome and fraught with points of clonal loss or contamination. Use of a robotic picker and gridder would alleviate these problems. Currently no such instrument exists in the greater Cleveland/Northeastern Ohio region. This robotic picking and gridding instrument will greatly enhance cDNA-arraying capabilities of the Laboratory of Gene Expression and the DNA Laboratory of Clinical Medicine CORE (DLCM) within the Lerner Research Institute of the Cleveland Clinic Foundation (CCF). The requested instrumentation will also be used to facilitate and advance research programs at institutions within the area (CCF and Case Western Reserve University). Major Users all have numerous NIH-funded research grants, which have component projects that can best, be conducted using the colony/phage picking and rearraying/cherry-picking or high density arraying features of this instrumentation system. All the Major User's projects require substantial needs to manipulate large phage or cDNA libraries. The Major Users projects are broad and include: I) identification of differentially expressed tumor suppressor genes or oncogenes in diseased tissue; 2) identification of interacting proteins with defined protein components in a complex biological protein mixture; 3) characterization of protein: protein interaction domains; and 4) identification of altered cellular signaling pathways and gene expression patterns in response to bioterroristic biological agents. The Q-bot system provides the versatility, resolution and capabilities to meet these diverse demands. In addition, the requested instrument system will be used to meet the needs of the occasional user. This service will be conducted within our existing Molecular Biotechnology Core, which provides support to investigators by performing protein sequencing, peptide synthesis, and related services. Due to the human genome project, the DNA arraying facilities have experienced greatly increased usage. Using the Q-bot, the DNA arraying facilities at CCF will be able to generate highly diverse types of arrays and maintain and replicate their valuable cDNA sequence verified 40K human and 15K mouse libraries. CCF has committed substantial resources to the developing the DNA Laboratory of Clinical Medicine and the Molecular Biotechnology Cores, including personnel to oversee its operation, and a fiscal plan to insure its operation and maintenance for the long-term. In sum, acquisition of the requested high resolution picking and gridding Q-bot system will have a significant impact on the progress and future development of biomedical research programs at multiple institutions in the region including CCF and CWRU.