High Throughput Microrepository for Genetic Materials

遗传物质高通量微存储库

基本信息

批准号：
8333400
负责人：
ROBERT C HAUSHALTER
金额：
$ 10.73万
依托单位：
PARALLEL SYNTHESIS TECHNOLOGIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8333400
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

项目摘要

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.

描述（由申请人提供）：遗传材料的高通量微存储库认识到迫切需要一种安全、快速和可靠的方法来保存和存档遗传样本和其他生物材料，例如合成 DNA 样本、独特的分离样本、质粒、法医样本NIH NCRR 最近召集的一个研究小组确定了最迫切需要的领域，并提出了几项建议。他们的主要建议清单中的第一项和第五项是“鼓励开发用于种质处理和冷冻保存的高通量和可扩展技术”和“支持将开辟新天地的新型‘高风险/高回报’保存技术”。我们通过遗传材料微存储库 (MGM) 技术直接成功地满足了这些需求，该技术提供完全可扩展的、非常高密度的样品存储，通过在光学复用编码珠阵列中提供样品制备-识别-存档-检索功能来自动检索。单一平台。在第一阶段的工作中，成功构建并测试了完全自动化的原型 MGM 平台。 MGM 包含密集的多孔玻璃或聚合物珠子、较小的珠子组或纸样，每个珠子都包含不同的 DNA 样本，这些样本已使用我们基于稀土的 Parallume 技术进行光学编码，从而可以对每个珠子进行光学识别独一无二。将 DNA 放置到任一单个大珠子上后，将珠子加载到珠子定位载玻片 (BLS) 中，将珠子光学隔离成平面单层以进行成像，并读取每个珠子的光学代码以确定该珠子的位置。特定的 DNA 样本。取出珠子并将其放置在所需位置，并通过 qPCR 确认处理过程中没有污染。通过进行额外的实验，包括采用新的改进的硬件设计并成功改变 Parallume 编码材料的化学成分以防止紫外线对 DNA 的损伤，我们已经解决了所有审稿人的担忧。我们现在已经构建并成功测试了功能齐全的原型 MGM 仪器，并且为了响应富有洞察力的审稿人的评论，我们修改了 Parallume 材料，以便允许使用更长波长的激发光 (365nm) 进行激发，这 (a) 防止光学解码样品过程中 DNA 的光化学分解，(b) 允许使用非紫外光学器件来聚焦光，(c) 允许使用更亮、寿命更长且更便宜的 LED 光源。一种新的表面化学物质将 DNA 牢固地结合到珠子上，直至其以化学方式释放，这将提供另一级别的样品污染保护。这里描述的改进的 MGM 代表了自该领域诞生以来在解决 DNA 和核酸快速扩展的存储需求方面取得的第一个实质性进展。采用光学编码珠技术的新存储范例提供了目前不可能实现的样本密度、自动化程度和简单性，并且具有可扩展性，可以在可预见的未来成功存储和检索任意数量的 DNA 样本。