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(申请人提供)：核酸越来越多地被用作许多领域的靶向分子标记，如生物恐怖检测、临床诊断、药物发现、生态和环境监测等生命科学领域。虽然实验室提取和纯化核酸的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为种类繁多的生物样本提供足够的通用性。此外，目前的大多数方法都不适用于非实验室环境。由于通常需要多个步骤、几件实验室设备和消耗品，因此这些程序的显著简化对于基于核酸的安全、通用和强大的应用是必要的。在这里，我们建议开发一种新的样品处理技术，可以在一步操作中实现核酸的提取和纯化。如上所述，这项技术可以立即成功地应用于许多领域。这一新工艺是基于使用高低压交替循环(压力循环技术或PCT)从样品中提取核酸的。通过控制加压下核酸与纯化基质的结合和解离，成功的PCT提取工艺将推广到核酸的纯化中。这个SBIR第一阶段的第一个目标是设计、制备和鉴定适合于在加压条件下纯化核酸的结合基质。与我们的合作者合作，我们将检查已知和新开发的矩阵，确定它们在高水平和低水平压力下的稳定性和亲和力。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，从而产生用于下游分析的产品。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，如杂交、序列扩增和定量。在SBIR第二阶段研究中，将进一步开发和优化可行的矩阵、原型仪器和测试模块，以实现商业化。 基于核酸的分析在现代生命科学中发挥着关键作用，如临床诊断、药物发现、法医学、环境监测和生物防御。随着人类基因组计划的完成和对生物王国基因组序列知识的爆炸性增长，从生物制品中快速提取和纯化核酸在改善公众健康和加快基础生物学研究方面变得越来越重要。毫无疑问，核酸的应用将继续扩大，并为人类带来天文数字的经济效益。为了促进基于核酸的安全、通用和强大的应用，我们建议开发一种基于高低压循环(压力循环技术或PCT)的新的样品处理技术。如果成功，这项技术可以立即应用于生命科学的许多领域。