High Throughput Microrepository for Genetic Materials

遗传物质高通量微存储库

• 批准号: 8058885
• 负责人: ROBERT C HAUSHALTER
• 金额: $ 46.35万
• 依托单位: PARALLEL SYNTHESIS TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8058885
• 关键词: Address; Ammonium; Archives; Area; Attenuated; Automation; Bacteria; Biocompatible Materials; Biodiversity; Biological Assay; Biological Preservation; Budgets; Buffers; Caliber; Chemicals; Chemistry; Code; Color; Computer software; Cryopreservation; DNA; DNA Binding; DNA Damage; DNA Markers; Data; Destinations; Detection; Development; Ensure; Fluorescence; Forensic Medicine; Future; Gel; Genetic; Genetic Materials; Glass; Government; Growth; Human Genome; Image; Image Analysis; Immobilization; Individual; Laboratories; Lasers; Life; Light; Liquid substance; Location; Microscope; Motion; National Center for Research Resources; Nucleic Acid Amplification Tests; Nucleic Acids; Optics; Organism; Output; Paper; Patients; Phase; Plants; Plasmids; Polymers; Preparation; Price; Printing; Process; Proteins; Protocols documentation; Published Comment; Radiation; Reader; Reading; Recommendation; Research; Resolution; Retrieval; Rice; Sampling; Scanning; Slide; Sorting - Cell Movement; Source; Speed; Staging; Surface; Suspension substance; Suspensions; System; Technology; Testing; Time; Tube; United States National Institutes of Health; University Hospitals; absorption; animal cloning; base; biobank; commercialization; density; design; fiberglass; functional genomics; high risk; human DNA; improved; instrument; monolayer; nanoparticle; novel; optical fiber; optical imaging; particle; prevent; prototype; repository; research study; response; software development; solid state; synthetic construct; ultraviolet damage

DESCRIPTION (provided by applicant): High Throughput Microrepository for Genetic Materials Recognizing the urgent need for a safe, rapid and reliable means of preserving and archiving genetic samples and other biological materials, such as synthetic DNA samples, unique isolated samples, plasmids, forensic samples, patient samples, samples to ensure future biodiversity in plants or animals, clones and archeological samples, a study panel recently convened by NIH NCRR identified the areas of most critical need and made several recommendations. The 1st and 5th items on their list of major recommendations were to "encourage the development of high throughput and scalable technologies for Germplasm processing and cryopreservation" and "support novel 'high risk/high return' preservation technologies that will break new ground". We have directly and successfully addressed these needs with our Microrepository for Genetic Materials (MGM) technology which affords completely scalable, very high density sample storage with automated retrieval from arrays of optically multiplexed encoded beads by providing sample preparation-identification-archiving-retrieval functions on a single platform. For the Phase I effort, a completely automated prototype MGM platform was successfully built and tested. The MGM contains dense arrays of porous glass or polymer beads, groups of smaller beads or paper swatches, each of which contains a different DNA sample, which have been optically encoded with our rare earth-based Parallume technology which allows each bead to be optically identified uniquely. After placement of the DNA onto either single large beads, the beads are loaded into Bead Localization Slides (BLS), which optically isolate the beads into a planar monolayer for imaging, and the optical code of each bead is read to determine the location of that particular DNA sample. The beads are retrieved and placed in the desired location and the lack of contamination during handling confirmed by qPCR. By performing additional experiments, including incorporation of new improved hardware designs and successfully changing the chemical composition of the Parallume encoding materials to prevent UV damage to the DNA, we have addressed all Reviewers' concerns. We have now built and successfully tested a completely functional prototype MGM instrument and, in response to the insightful Reviewers' comments, we have modified the Parallume materials so as to allow excitation with a longer wavelength of excitation light (365nm) which (a) prevents the photochemical decomposition of the DNA during optically decoding the samples, (b) allows non-UV optics to be used to focus the light and (c) lets much brighter, longer lived and less expensive LED light sources be used. A new surface chemistry, which strongly binds the DNA to the bead until it is chemically released, will provide yet another level of sample contamination protection. The improved MGM described here represents the first substantial progress since inception of the field in addressing the rapidly expanding storage requirements for DNA and nucleic acids. A new storage paradigm which employs optically encoded bead technology provides a sample density, degree of automation and simplicity not currently possible and with scalability to successfully store and retrieve any number of DNA samples for the foreseeable future. PUBLIC HEALTH RELEVANCE: In recent years there has been a very large increase in the number of samples of genetic materials such as DNA that have been synthesized or isolated from organisms. Some of these samples, such as an individual's DNA, forensic samples or genetic material from a new strain of bacteria are one-of-a-kind and cannot be replaced. However, there is currently no means to organize or automatically archive and retrieve DNA samples in an automatic and high throughput manner. Because of the increase in throughput from laboratories, governments, hospitals and universities, the problem is only expected to grow in the future. We offer here a design of a new instrument platform designated as the Microrepository for Genetic Materials (MGM). The samples will be stored in arrays of porous glass beads with each of the beads displaying an optical code or signature that uniquely identifies that particular sample and the DNA within that bead. The DNA samples are each placed into a single highly porous bead with a diameter of 50 microns (0.05mm). The beads are optically encoded with our ParallumeTM technology which allows each bead to be identified by the intensity ratios of the different colors emitted by the bead upon excitation. Thus, thousands of beads can be mixed together, each identified by its unique optical code and removed from the storage array using an optical fiber to pick it up. After checking the optical signature by examining the emitted light when the bead is picked up by the optical fiber, the bead can be placed in the desired destination location. Thus, we are able to design and build a repository that can store hundreds of thousands of DNA samples in a robust and inexpensive manner. This MGM technology is completely scalable and should provide rapid and accurate DNA storage and retrieval capacity for the foreseeable future.

描述（由申请人提供）：高通量微孔位用于遗传材料，以确认迫切需要安全，快速和可靠的方法来保存和存档遗传样品以及其他生物学材料，例如合成DNA样品，独特的孤立样品，质粒，质粒，法医样品，患者样品，样品，以确保植物或动物，克隆和考古样本中未来的生物多样性，NIH NCRR最近召集的研究小组确定了最迫切需要的领域，并提出了一些建议。他们的主要建议清单上的第一和第五项是“鼓励开发高吞吐量和可扩展的技术来用于种质处理和冷冻保存”，并“支持新型的“高风险/高回报”保存技术，这些技术将破坏新地面”。我们已经通过微验证机构（MGM）技术直接成功地满足了这些需求，该技术提供了完全可扩展的，非常高密度的样品存储，并通过提供样品制备识别识别 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构 - 架构识别函数一个平台。在第一阶段的工作中，成功构建和测试了一个完全自动化的原型MGM平台。米高梅包含多孔玻璃或聚合物珠的密集阵列，较小的珠子或纸色板组，每个珠子包含不同的DNA样品，这些样品已通过我们的稀土基本派别在光学上编码，允许每个珠子被光学地识别独特。将DNA放置在任何一个大珠上后，将珠子加载到珠子定位载玻片（BLS）中，将珠子光学地隔离到平面单层中进行成像，并读取每个珠子的光学代码以确定其位置特定的DNA样品。珠子被检索并放置在所需的位置，并且在QPCR确认的处理过程中缺乏污染。通过进行其他实验，包括合并新的改进的硬件设计，并成功地更改andallume编码材料的化学组成，以防止对DNA的紫外线损害，我们已经解决了所有审阅者的关注点。现在，我们已经构建并成功地测试了一种完全功能的MGM仪器，并响应有见地的审稿人的评论，我们修改了阶梯材料，以便使用更长的激发光（365nm）允许激发，这（a）可以预防。 DNA在光学解码样品过程中的光化学分解，（b）允许使用非UV光学器件来聚焦光线，并且（c）可以使用更明亮，更长的寿命和较便宜的LED光源。一种新的表面化学物质将DNA强烈结合到化学释放，直到其化学释放为止，它将提供另一个水平的样品污染保护。此处描述的改进的MGM代表了自田间以来的第一个实质性进展，以解决DNA和核酸迅速扩展的储存要求。采用光学编码珠技术的新存储范式提供了当前不可能的样品密度，自动化程度和简单性，并且具有可扩展性，可在可预见的将来成功存储和检索任何数量的DNA样品。 公共卫生相关性：近年来，遗传材料的样本数量（例如已合成或与生物体中分离出来的DNA）的数量大大增加。这些样品中的一些，例如个体的DNA，法医样品或来自新细菌菌株的遗传物质是一种独一无二的，无法替代。但是，目前没有手段以自动且高通量的方式组织或自动存档并检索DNA样品。由于实验室，政府，医院和大学的吞吐量有所增加，因此该问题只会在未来增长。 我们在这里提供了一个新的仪器平台的设计，该平台被指定为遗传材料（米高梅）的微孔道。样品将存储在多孔玻璃珠的阵列中，每个珠子显示光学代码或签名，这些光学代码或签名独特地识别该珠子中的特定样品和DNA。将DNA样品分别放入直径为50微米（0.05mm）的单个高度多孔珠中。这些珠子是通过我们的parallumetm技术在光学上编码的，该技术使每个珠子都可以通过激发后珠子发出的不同颜色的强度比识别。因此，可以将数千个珠子混合在一起，每个珠子都通过其唯一的光学代码识别，并使用光纤从存储阵列中删除。通过检查光纤拾取珠子时检查光学签名后，可以将珠子放置在所需的目的地位置。因此，我们能够设计和建造一个存储库，该存储库可以以健壮且廉价的方式存储数十万个DNA样品。这种米高梅技术是完全可扩展的，应在可预见的未来提供快速准确的DNA存储和检索能力。