Automated Large and Mid Scale Nucleic Acid Preparation

描述（由申请人提供）：本研究涉及开发一次性纯化盒和台式电泳处理仪器，可在60分钟内从重组培养物中纯化1毫克或更多的质粒DNA。在这项工作中采用的纯化方法是经过验证的电泳分离技术的衍生物，该技术是我们公司开发的用于自动迷你准备DNA纯化的技术。第一阶段之前获得的数据表明，从直接装入原型盒的50毫升液体细菌培养物中可以自动纯化高达0.3毫克的高纯度质粒DNA。这些初步数据表明，盒式培养基可以处理多达150毫升的培养物，从而产生1毫克或更多的质粒DNA。第一阶段的工作将建立产品的概念验证。第一阶段的目标包括：优化盒式设计，以确定放大过程的局限性，优化电泳电压程序，以减少运行时间到一个小时，没有产量损失，并以最少的用户步骤。该方法还将用于从细菌和哺乳动物细胞中纯化基因组DNA。第一阶段将建造一个微处理器控制电路，它将成为第二阶段将开发的实际处理仪器的基础。这项研究的产品将比目前可用的任何仪器或手动套件更快，更容易使用。该产品的一次性产品预计每个样品的成本为10美元，而台式仪器的价格约为2000美元。该方法基本上没有活动部件。这项工作将大大减少提纯质粒DNA所需的人力和成本。质粒和基因组DNA的自动纯化参与了数亿美元的核酸纯化市场。该产品每年有可能产生1000万至2000万美元的收入。

DESCRIPTION (provided by applicant): This research involves the development a disposable purification cassette and bench-top electrophoretic processing instrument that will purify one milligram or more of plasmid DNA from recombinant culture in 60 minutes. The purification method to be employed in this work is a derivative of proven electrophoretic separation technology that our company developed for automated mini-prep DNA purification. Data obtained prior to Phase I demonstrated that up to 0.3 milligrams of highly pure plasmid DNA could be automatically purified from 50 mls of liquid bacterial culture loaded directly into a prototype cassette. This preliminary data indicates that cassettes could be made to process up to 150 mls of culture and thus yield 1 mg or more of plasmid DNA. Phase I work will establish the proof-of-concept for the product. Aims of Phase I include: optimization of the cassette design to determine the limitations of the scaled-up process, and optimization of the electrophoretic voltage program to reduce the run time to one hour, without yield loss, and with minimal user steps. The process will also be tested for purification of genomic DNA from bacterial and mammalian cells. A microprocessor control circuit will be constructed in Phase I that will be the basis of the actual processing instrument to be developed in Phase II. The products from this research will be faster and far easier to use than any instrument or manual kit currently available. The disposables of this product are projected to cost $10 per sample, while the bench-top instrument will have a price of ~$2000. The method has essentially no moving parts. This work will greatly reduce the labor and cost required to purify milligram amounts of plasmid DNA. Automated purification of plasmid and genomic DNA participates in a multi- hundred million-dollar nucleic acid purification market. The product has the potential to generate $10-20 million per year in revenue. PUBLIC HEALTH RELEVANCE: The work will create a bench-top instrument and disposable cassettes that will allow for fully automated purification of milligram quantities of plasmid and genomic DNA. This technology will be significantly less expensive and far less time consumptive that any existing instrument or manual kit available.