Materials for Isolation of Nucleic Acids from Whole Blood without a Lysis Step

描述（由申请人提供）：该二期项目继续开发和应用新材料，特别是用于直接从细胞、组织和生物流体（包括全血）中快速分离DNA的磁性微粒。该材料消除了执行裂解步骤和从复杂材料中捕获DNA的需要，从而简化了样品制备方法。用于结合和释放核酸的功能化微粒利用专有化学，使它们能够在几乎任何生物条件下自发和快速地裂解细胞并捕获暴露的DNA。因此，DNA可以从全血中分离出来，而无需任何裂解步骤。独特的表面磷基团化学使核酸以特殊的强度结合，直到通过独特的裂解机制释放，其中连接基团被简单的化学反应选择性地破坏，以便从固体表面释放DNA。化学裂解释放而不是传统的洗脱，可以防止过早的、不受控制的物质释放，并产生纯核酸。分离的DNA与下游应用如PCR兼容。正在探索其他连接基团和裂解方法。第二阶段将发展完成完整合成的专业知识，包括制备磁芯、涂覆磁芯以及用连接剂和头基进行功能化。大规模制造功能化磁性粒子的能力将得到发展。我们制作了一个从全血中分离DNA的原型试剂盒。我们认为这些新材料的应用范围正在扩大到质粒纯化，从病毒中分离DNA，以及从尿液，食品和培养基中的感染性微生物中捕获DNA，而无需进一步分离步骤。这些材料有望在广泛的应用中简化样品制备和DNA提取。该二期项目继续开发和应用新材料，特别是用于从细胞、组织和生物流体（包括全血）中以极快的方式直接分离DNA的磁性微粒。该材料消除了执行裂解步骤和从复杂基质中捕获DNA的需要，从而简化了样品制备方法。这些材料将在生命科学和诊断行业的大量应用中找到用途，其中DNA提取是必要的。

DESCRIPTION (provided by applicant): This Phase II project continues the development and application of new materials, especially magnetic microparticles for directly isolating DNA from cells, tissues, and biological fluids, including whole blood, in an extremely rapid format. The materials eliminate the need to perform a lysis step and capture DNA from complex materials thereby simplifying sample preparation methods. The functionalized microparticles for binding and releasing nucleic acids utilize a proprietary chemistry which enables them to spontaneously and rapidly lyse cells and capture the exposed DNA in situ under virtually any biological conditions. As a result DNA can be isolated from whole blood without any lysis step. The unique surface phosphonium group chemistry causes nucleic acids to bind with exceptional strength until released by a unique cleavage mechanism where a linker group is selectively broken by a simple chemical reaction in order to free DNA from the solid surface. Release by chemical cleavage rather than traditional elution prevents premature, uncontrolled release of material and yields pure nucleic acid. Isolated DNA is compatible with downstream applications such as PCR. Additional linking groups and cleavage methods are being explored. Phase II will develop the expertise to perform the complete synthesis including preparing the magnetic cores, coating the cores, and functionalizing with linker and head groups. The capability to manufacture the fuctionalized magnetic particles on a large scale will be developed. We have produced a prototype kit for isolating DNA from whole blood. We areing the range of applications of these new materials is being expanded to plasmid purification, isolation of DNA from viruses, and capture of DNA from infectious microorganisms in urine, foodstuffs and culture media with no further isolation steps. The materials are expected to streamline sample preparation and DNA extraction across a broad variety of applications. This Phase II project continues the development and application of new materials, especially magnetic microparticles for directly isolating DNA from cells, tissues, and biological fluids, including whole blood, in an extremely rapid format. The materials eliminate the need to perform a lysis step and capture DNA from complex matrices thereby simplifying sample preparation methods. The materials will find use in a great number of applications in the life sciences and diagnostics industries for which DNA extraction is necessary.