Development and Implimentation of a Materials Repository for the PSI

描述(由申请人提供)： 集中获取关键生物试剂，如质粒克隆，有望促进科学和我们对生物学的理解。这些资源越能被公开分享，它们就越能推动研究。在蛋白质结构研究过程中，蛋白质结构倡议(PSI)已经产生了数千个有用的蛋白质表达克隆，他们计划通过拟议的PSI-材料库与社区共享。哈佛蛋白质组学研究所(HIP)在质粒的生产、序列验证和使用方面拥有五年多的经验，包括生物防御相关生物和数千个人类基因的基因组规模收集。HIP正在进行的工作包括在哺乳动物细胞中进行高通量(HT)cDNA过表达和shRNA击倒分析，在酵母中进行过表达分析，HT蛋白纯化，以及蛋白质微阵列的生产和使用。因此，我们深入了解克隆用户的需求，因为使用克隆是我们研究的核心。 从一开始，HIP就一直不受限制地共享它产生的克隆，我们创建了一个复杂的基础设施来存储、维护和分发克隆到世界各地。最近，我们已经开始与其他希望通过我们分发克隆的实验室共享这一基础设施。HIP一直是这一领域的创新者，为克隆生产和管理建立了全自动化的标准操作规程，包括机器人进行常规条形码检查，发明了一种新的自动化HT菌落挑选方法，开发了低成本和高质量的HT DMA微型制备剂，并开发了企业级软件来管理从生产和序列验证到维护和分发的克隆数据。HIP与ThermoElectron一起帮助开发了首个全自动-80C克隆存储系统，该系统使用专门的2D条形码跟踪160,000个管子，有望彻底改变生物材料的存储和分配。 克隆分发的坚实基础设施的存在，再加上哈佛大学作为一个机构的稳定性，为整合来自PSI站点的克隆提供了一个理想的机会。我们建议创建一个独特的用户友好的PSI专用门户网站，供用户检查、查询和订购PSI克隆。该门户将重点关注与PSI克隆集相关的独特用户需求。该建议建立在我们目前成功的简化MTA流程以及我们精心开发的机制的基础上，以确保准确的克隆跟踪、可靠的克隆生存能力和强大的备份程序。摘要：我们建议使用软件开发和自动物理存储设备来创建DMA分子的储存库，科学家可以用它来研究许多蛋白质及其在疾病中的作用。这个存储库将向任何从事学术研究的人开放，并将极大地加快发现速度。

DESCRIPTION (provided by applicant): Centralized access to key biological reagents, such as plasmid clones, promises to accelerate science and our understanding of biology. The more these resources are openly shared, the more they will drive research. In the course of protein structural studies, the Protein Structure Initiative (PSI) has produced thousands of useful protein expression clones that they plan to share with the community through a proposed PSI-Materials Repository. The Harvard Institute of Proteomics (HIP) has more than five years of experience in the production, sequence verification, and use of plasmids, including genome scale collections for biodefense related organisms and thousands of human genes. Ongoing work at HIP includes highthroughput (HT) cDNA over-expression and shRNA knock-down assays in mammalian cells, overexpression assays in yeast, HT protein purification, and the production and use of protein microarrays. Thus, we understand deeply the needs of clone users, because using clones is central to our research. From its beginning, HIP has always shared the clones it has produced without restriction and we have created a sophisticated infrastructure to store, maintain, and distribute clones worldwide. More recently, we have begun sharing this infrastructure with other labs who would like to distribute their clones through us. HIP has been an innovator in this area, establishing fully automated SOPs for clone production and management that include routine barcode checks by robots, invention of a novel automated HT colony picking approach, development of low cost and high quality HT DMA miniprep chemistries, and development of enterprise class software that manages clone data from production and sequence validation to maintenance and distribution. Together with ThermoElectron, HIP has helped to develop and now owns a first-of-its-kind fully automated -80¿C clone storage system that tracks 160,000 individual tubes with specialized 2D barcodes, which promises to revolutionize the storage and distribution of biomaterials. The existence of this solid infrastructure for clone distribution, coupled with Harvard's stability as an institution, offer an ideal opportunity to incorporate the clones from the PSI sites. We propose to create a unique user-friendly PSI-specific web portal for users to examine, query and order the PSI clones. This portal will focus on the unique user needs related to the PSI clone set. The proposal builds on our currently successful simplified MTA process, and our carefully developed mechanisms to ensure accurate clone tracking, reliable clone viability and robust back up procedures. Lay Abstract: We propose to use software development and an automated physical storage device to create a reposity of DMA moleucles that scientists can use to study many proteins and their roles in disease. This repository will be available to anyone doing academic research and will dramatically speed discovery.