Nucleic Acid Purification Using Pressure Cycling Technology

描述（申请人提供）：核酸作为靶向分子标记越来越多地应用于许多领域，如生物恐怖检测、临床诊断、药物发现、生态和环境监测以及生命科学的其他领域。虽然核酸提取和纯化的实验室方法组合通常可以满足研究科学家的需要，但目前没有一种方法能够为各种各样的生物样品提供足够的多功能性。此外，目前大多数方法不适合非实验室环境。由于通常需要多个步骤，几件实验室设备和消耗品，因此为了安全，通用和稳健的基于核酸的应用，需要显著简化这些程序。在这里，我们建议开发一种新的样品处理技术，将实现核酸提取和纯化在一个操作步骤。如上所述，这项技术可以立即成功地应用于许多领域。这种新方法是基于使用高压和低压交替循环（压力循环技术或PCT）从样品中提取核酸。通过在压力下控制核酸与纯化基质的结合和解离，成功的PCT提取过程将花费在核酸的纯化上。SBIR一期的第一个目标是设计、制备和鉴定适合在压力条件下纯化核酸的结合基质。与我们的合作者合作，我们将检查已知的和新开发的矩阵，确定它们的稳定性和亲和力，暴露在高和低水平的压力循环下。第二个目标是设计和制造原型模块和设备，允许一步去除污染分子和纯化核酸，从而产生下游分析的产品。使用PCT过程获得的DNA的数量和质量将使用现代核酸分析方法进行测试，例如杂交，序列扩增和定量。可行的矩阵、原型仪器和测试模块将在SBIR二期研究中进一步开发和优化商业化。

DESCRIPTION (provided by applicant): Nucleic acids are increasingly used as targeted molecular markers in many fields, such as bioterror detection, clinical diagnostics, drug discovery, ecological and environmental monitoring, and other areas in life sciences. Although a combination of laboratory methods for nucleic acid extraction and purification can usually meet the needs of the research scientist, no single method currently offers sufficient versatility for the wide variety of biological samples. In addition, most current methods are not suitable for the non-laboratory environment. Since multiple steps, several pieces of laboratory equipment and consumables are typically required, significant simplification of these procedures are necessary for safe, versatile, and robust nucleic acid based applications. Here, we propose to develop a novel sample processing technology that will achieve nucleic acid extraction and purification in a single operational step. This technology could be immediately and successfully employed in many fields as mentioned above. The novel process is based on the use of cycles of alternating high and low pressure (Pressure Cycling Technology or PCT) to extract nucleic acids from samples. By controlling the binding and dissociation of nucleic acids to purification matrices under pressure, the successful PCT extraction process is to be expended in the purification of nucleic acids. The first goal of this SBIR Phase I is to design, prepare and identify binding matrices that are suitable for purification of nucleic acids under pressure conditions. Working with our collaborators, we will examine known and newly developed matrices, determine their stability and affinity, under exposure to cycles of high and low levels of pressure. The second goal is to design and fabricate prototype modules and equipment that will allow single-step removal of contaminating molecules and purification of nucleic acids, thus yielding products for the downstream analysis. The quantity and quality of DNA obtained using the PCT process will be tested using modern nucleic acid analytical methods, such as hybridization, sequence amplification and quantification. Feasible matrices, prototype instruments, and test modules will be further developed and optimized for commercialization in an SBIR Phase II study. Nucleic acid based analyses play key rolls in modern life sciences, such as clinical diagnostics, drug discovery, forensics, environmental monitoring and biodefense. By the completion of human genome project and undertaking the explosive expansion in knowledge of genomic sequences of the biological kingdom, rapid extraction and purification of nucleic acids from biologicals become increasingly important in improving public health and accelerate basic biological research. There is no doubt that nucleic acid applications will continue to expand and produce astronomical economic benefits to human being. To facilitate safe, versatile, and robust nucleic acid based applications, we propose to develop a novel sample processing technology based on the use of cycles of high and low pressure (Pressure Cycling Technology or PCT). If successful, this technology could be immediately employed in many fields of life sciences.