Rapid, Very Low Cost, Automated DNA Purification Device

快速、成本极低的自动化 DNA 纯化装置

• 批准号: 8538462
• 负责人: WILLIAM P MACCONNELL
• 金额: $ 48.31万
• 依托单位: MACCONNELL RESEARCH CORPORATION
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-15 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8538462
• 关键词: Bacteria; Bathing; Blood; Blood Volume; Buffers; Businesses; Cells; Chemistry; Clinical; Computer software; Contracts; Cytolysis; DNA; DNA purification; Detection; Devices; Diagnostic Procedure; Gel; Genomics; Injection of therapeutic agent; Instruction; Laboratories; Left; Mammalian Cell; Manuals; Marketing; Methodology; Methods; Molds; Molecular Biology; Nucleic Acids; Phase; Plants; Plastics; Power Sources; Preparation; Process; Protocols documentation; Recovery; Research; Reverse Transcriptase Polymerase Chain Reaction; Running; Saliva; Sampling; Sepharose; Small Business Innovation Research Grant; Technology; Time; Tissues; Variant; Whole Blood; Work; cost; design; improved; innovation; instrument; new technology; next generation sequencing; plasmid DNA; platinum electrode; programs; prototype; touchpad; voltage

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.

描述（由申请人提供）：这项工作将创造创新的产品，允许快速，低成本，自动化，基因组DNA纯化从几个样品类型和浓度。该新技术使用一次性盒装置和低成本、可重复使用的处理支架，通过独立的半干式电泳方法自动纯化DNA。Cascadium可以被构建为包含多个泳道，其允许在一次运行中同时纯化几个或多达96个样品。该方法不需要移动部件，并且可以在不到10分钟的时间内对某些样品类型进行操作。每个样本的盒的预计成本为0.4 - 0.8美元，而便携式处理支架的成本将低于1-12样本版本的250美元，或96样本版本的750美元。该产品如果完全实现，可能会彻底改变临床或参考实验室诊断程序的DNA制备方式。它将降低成本，节省劳动力和材料。 在第一阶段进行的纯化盒和处理装置原型的工作表明，从血液、细菌、唾液或组织中制备的DNA是高纯度的，可以直接用于PCR扩增以及其他分子生物学应用。第一阶段的工作还表明，该方法可以开始与宽范围的样品浓度，因为起始样品可以稀释100-500倍，和方法仍然产生活性DNA模板。 该纯化方法利用了我们公司开发的自动质粒DNA纯化技术的变体，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试剂卡和处理支架仪器的工作原型。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)确定纯化DNA的活性范围。5)制定小体积血液的方案。6)研制12孔和96孔工艺摇架仪器。7)完成2孔、6孔、12孔和24孔试剂卡设计，并签订注塑模具建造合同。8)起草说明书和产品规格。我们公司将能够在第二阶段之后直接制造和销售从这项工作中开发的仪器和卡匣。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。