Rapid, Very Low Cost, Automated DNA Purification Device

描述（由申请人提供）：这项工作将创造创新的产品，允许从几种样品类型和浓度中快速，低成本，自动化的基因组DNA纯化。这项新技术使用一次性卡式装置和低成本、可重复使用的处理支架，通过独立的半干电泳方法自动纯化DNA。盒式磁带可以构建成包含多个通道，允许同时净化几个，或在一次运行中多达96个样品。该方法不需要移动部件，并且可以在不到10分钟的时间内完成某些样品类型。盒式磁带的预计成本为每个样品0.4 - 0.8美元，而便携式处理支架的1-12个样品版本的成本不到250美元，或96个样品版本的成本不到750美元。这个产品，如果完全实现，可能会彻底改变DNA为临床或参考实验室诊断程序准备的方式。它将降低成本，节省人工和材料。第一阶段使用纯化盒原型和处理设备进行的工作表明，从血液、细菌、唾液或组织中制备的DNA纯度很高，可直接用于PCR扩增以及其他分子生物学应用。第一阶段的工作还表明，该方法可以在很宽的样品浓度范围内开始，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开发的用于自动质粒DNA纯化的技术变体，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加工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程序。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纯化DNA。3)改进裂解化学。4)测定纯化DNA的活性范围。5)制定小容量血液的方案。6)开发12井和96井加工摇床仪表。7)完成2井、6井、12井和24井盒的设计和合同，以便建造注塑模具。8)起草使用说明书和产品规格书。我们公司将能够生产和销售从这项工作开发的仪器和磁带直接二期后。这些产品的市场潜力估计约为5000万美元。该产品也适用于临床实验室和可能的医生办公室设置。

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.