Rapid, Very Low Cost, Automated DNA Purification Device

描述(申请人提供)：这项工作将创造创新的产品，允许快速，低成本，自动化，从几种类型和浓度的样本中提纯基因组DNA。这项新技术使用一次性盒式装置和低成本、可重复使用的处理摇篮，通过独立的半干凝胶电泳法自动纯化DNA。盒可以构造为包含多个通道，允许在一次运行中同时提纯几个或最多96个样品。该方法不需要移动部件，可以在不到10分钟的时间内完成某些样品类型的测试。预计盒式磁带的成本为每个样本0.4-0.8美元，而便携式处理支架的1-12个样本版本的成本不到250美元，或96个样本版本的750美元。如果完全实现这种产品，可能会彻底改变DNA准备用于临床或参考实验室诊断程序的方式。这将降低成本，节省人力和材料。在第一阶段使用净化盒和处理设备的原型进行的工作表明，从血液、细菌、唾液或组织中提取的DNA纯度高，可直接用于聚合酶链式反应扩增和其他分子生物学应用。第一阶段的工作还表明，该方法可以从很大范围的样品浓度开始，因为起始样品可以稀释100-500倍，并且该方法仍然可以产生活性DNA模板。该纯化方法利用了我们公司为自动纯化质粒DNA而开发的各种技术，并结合了无需运行缓冲液的琼脂糖凝胶运行技术。第二阶段的工作将产生盒式磁带和处理摇篮仪器的工作原型。第二阶段的目标是：1)进一步优化电泳电压方案。2)提高细菌和哺乳动物细胞的检测下限，包括从单个哺乳动物细胞中提纯DNA。3)改进裂解化学。4)确定纯化DNA的活性范围。5)制定一项适用于少量血液的方案。6)研制12井、96井加工摇篮仪器。7)最终确定2、6、12和24孔盒的设计和注射模建造合同。8)起草使用说明书和产品说明书。我们公司将能够在第二阶段之后直接制造和销售通过这项工作开发的仪器和盒式磁带。这些产品的市场潜力估计为5000万美元。该产品还适用于临床实验室，可能还适用于医生的办公室环境。 与公共健康相关：这项提议将创造创新的产品，允许快速、低成本、自动化、多样本地从多种样本类型和浓度中提纯基因组DNA。这项新技术使用一次性、可回收的盒式装置和低成本的处理摇篮，通过电泳手段自动提纯DNA，而不需要运行缓冲池或铂电极。

DESCRIPTION (provided by applicant): This work will create innovative products that allow rapid, low cost, automated, genomic DNA purification from several sample types and concentrations. This novel technology uses a disposable cassette device and low cost, reusable processing cradle to automatically purify DNA by self-contained, semi-dry electrophoretic methodology. Cassettes can be constructed to contain multiple lanes that allow simultaneous purification of a few, or up to 96 samples in one run. The method requires no moving parts and can be performed, with some sample types, in less than 10 minutes. The projected cost of cassettes is $0.4 - $0.8 per sample, while the portable processing cradle will cost less than $250 for a 1-12 sample version, or $750 for a 96-sample version. This product, if fully realized, could revolutionize the way that DNA is prepared for clinical or reference laboratory diagnostic procedures. It will reduce costs and save labor and materials. Work performed in Phase I with prototypes of the purification cassette and processing devices showed that DNA prepared from blood, bacteria, saliva or tissue is highly pure and can be used directly in PCR amplification, as well as other molecular biology applications. Phase I work also showed that the method could begin with a wide range of sample concentrations, as the starting sample could be diluted 100-500 fold, and method still yielded active DNA template. The purification method utilizes a variation of technology that our company developed for automated plasmid DNA purification, combined with technology for running agarose gels without running buffer. Phase II work will result in the working prototypes for the cassettes and processing cradle instruments. Aims of Phase II are to: 1) Further optimize the electrophoretic voltage programs. 2) Improve the lower detection limit of bacterial and mammalian cells, including purification of DNA from single mammalian cells. 3) Improve the lysis chemistry. 4) Determine the scope of activity of the purified DNA. 5) Develop a protocol for small volumes of blood. 6) Develop the 12 and 96-well processing cradle instruments. 7) Finalize the 2, 6, 12 and 24-well cassette design and contract for the construction of injection molds. 8) Draft an instruction manual and product specifications. Our company will be able to manufacture and sell instruments and cassettes developed from this work directly after Phase II. The market potential for these products is estimated to be ~$50 million. The products are also applicable to clinical labs and possibly doctor's office settings. PUBLIC HEALTH RELEVANCE: This proposal will create innovative products that allow rapid, low cost, automated, multi-sample genomic DNA purification from many sample types and concentrations. The novel technology uses disposable, recyclable cassette device and low cost processing cradle to automatically purify DNA by electrophoretic means, without running buffer baths or platinum electrodes.