Nucleic Acid Purification Using Pressure Cycling Technology

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.