Nucleic Acid Purification Using Pressure Cycling Technology

使用压力循环技术纯化核酸

• 批准号: 7225657
• 负责人: Feng Tao
• 金额: $ 15万
• 依托单位: PRESSURE BIOSCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-05 至 2008-09-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7225657
• 关键词: Affinity; Area; Bacteriophage lambda; Binding; Biological; Biological Assay; Biological Markers; Biological Sciences; Buffers; Charge; Chemistry; Class; Clinical; Condition; Coupled; Crude Extracts; DNA; Detection; Detergents; Development; Diagnostic; Dissociation; Economics; Electric Wiring; Engineering; Environment; Environmental Monitoring; Equipment; Evaluation; Excision; Exposure to; Extravasation; Feasibility Studies; Fluorescence; Forensic Medicine; Genomics; Glass; Goals; Human; Human Genome Project; Ion Exchange; Knowledge; Laboratories; Liquid substance; Manuals; Mediating; Methods; Monitor; Nucleic Acid Binding; Nucleic Acids; Numbers; Performance; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Play; Polychloroterphenyl Compounds; Polymers; Preparation; Prevention, Clinical, and Therapeutic Subcommittee; Procedures; Process; Protein-Bound Iodine Tests; Public Health; Recovery; Research; Robotics; Sampling; Scheme; Scientist; Shock; Silicon Dioxide; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Solid; Solutions; Source; System; Technology; Testing; Uncertainty; Work; analytical method; base; biodefense; biological research; commercialization; design; drug discovery; high throughput analysis; improved; instrument; miniaturize; novel; nucleic acid purification; particle; pressure; prototype; success

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.

描述（由申请人提供）：核酸越来越多地用作许多领域的靶向分子标记，例如生物疗法检测，临床诊断，药物发现，生态和环境监测以及生命科学领域的其他领域。尽管用于核酸提取和纯化的实验室方法的组合通常可以满足研究科学家的需求，但目前尚无单一方法为各种生物样品提供足够的多功能性。另外，大多数当前方法不适合非实验室环境。由于多个步骤，通常需要几种实验室设备和消耗品，因此对于安全，多功能且基于核酸的核心应用应用是必需的这些程序的大量简化。在这里，我们建议开发一种新型的样品处理技术，该技术将在单个操作步骤中实现核酸提取和纯化。如上所述，该技术可以立即并成功地用于许多领域。新过程基于使用高压和低压（压力循环技术或PCT）的循环从样品中提取核酸。通过控制核酸在压力下与纯化基质的结合和解离，可以在核酸的纯化中进行成功的PCT提取过程。该SBIR I期的第一个目标是设计，准备和识别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，我们将研究已知的和新开发的矩阵，确定其稳定性和亲和力，并在暴露于高水平和低水平压力的周期中。第二个目标是设计和制造原型模块和设备，这些模块和设备将允许单步去除污染分子并纯化核酸，从而产生产物进行下游分析。使用现代核酸分析方法（例如杂交，序列扩增和定量）测试使用PCT过程获得的DNA的数量和质量。在SBIR II期研究中，将进一步开发并优化可行的矩阵，原型仪器和测试模块。 基于核酸的分析在现代生命科学中发挥关键作用，例如临床诊断，药物发现，取证，环境监测和生物陷入困境。通过完成人类基因组项目并在生物界的基因组序列知识中进行爆炸性扩张，从生物学中快速提取和纯化核酸在改善公共卫生和加速基本生物学研究方面越来越重要。毫无疑问，核酸的应用将继续扩展并为人类带来天文经济利益。为了促进基于核酸的安全，多功能和鲁棒的应用，我们建议根据使用高压和低压循环（压力循环技术或PCT）开发一种新型的样品处理技术。如果成功的话，这项技术可以立即在生命科学领域的许多领域使用。