NANODROP AND PCR USAGE

Nanodrop 和 PCR 用途

• 批准号: 7953866
• 负责人: PETER JS SMITH
• 金额: $ 1.12万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7953866
• 关键词: Ants; Atlantic Ocean; Biochemical; Centrosome; Cloning; Computer Retrieval of Information on Scientific Projects Database; DNA; Data; Detection; Development; Environment; Equipment; Evolution; Fellowship; Funding; Gene Expression; Genetic Processes; Genome; Goals; Grant; Human; Individual; Institution; Invertebrates; Knowledge; Laboratories; Life; Life Style; Microbe; Molecular; Nematoda; Nucleic Acids; Nutritional status; Pattern; Physiological; Population Genetics; Preparation; Proteins; RNA; Research; Research Personnel; Resources; Rivers; Role; Sampling; Science; Scientist; Services; Source; Staging; System; Time; United States National Institutes of Health; Variant; Wolbachia; Woman; Work; density; functional genomics; genome sequencing; member; microbial; mutualism; population survey; protocol development; technology development

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This equipment is utilized by local scientists at MBL in a service capacity. Individual brief research descriptions follow: Sogin: Our focus is on centrosome-associated RNAs and their possible role in the formation and function of centrosomes. The ability to quantitate nucleic acids and proteins in our biochemical preparations is essential to our research. The nanodrop provides this capability using minute quantities of sample and with high sensitivity. Our research is currently funded by NIH GM075163 (Cloning and analysis of centrosome-associated RNA). Huber: The crustal biosphere remains under-sampled, and our knowledge of what microbes are present and how they are distributed in this dynamic geochemical environment over time and space is fragmentary. We will use the DNA Engine Opticon Real-Time PCR Detection System from Bio-Rad to quantify microbial groups in subseafloor samples from throughout the Pacific (and eventually Atlantic) Ocean. We do not have the resources to carry out quantitative PCR in the Bay Paul Center. Members of the Bordenstein and Wernergreen laboratories have successfully used the BioCurrents machine, and we will use their expertise in our own protocol development. Funding for development of this technology will be provided by Dr. Huber's L'Oreal Fellowship For Women In Science and start-up package provided by the MBL Director, Dr. Gary Borisy. If additional grants are used, we will let you know. Wernergreen: This study explores the effects of bacterial lifestyle (free-living versus host-associated) on rates and patterns of genome evolution and population genetic processes. The project includes obtaining new genome sequence data and surveying population-level variation for select endosymbiotic species. We rely on the nanodrop to determine quality and concentration of DNA at several steps during our molecular work. Grant ID: 2R01GM062626-06A1 Wernergreen: This project integrates ecological, physiological, and functional genomic approaches to examine mechanisms of plasticity in an obligate bacterial mutualism. Our central goal is to understand how endosymbionts of ants respond to variation at the level of the host. We use the qPCR machine to quantify bacterial densities and gene expression patterns in hosts of distinct developmental stages and nutritional status. Bordenstein: We used the Nanodrop over the past year to quantify DNA and RNA concentrations of Wolbachia pipientis, a bacterial symbiont of invertebrates that also causes elephiantiasis and river blindenss in humans infected by filarial nematodes that harbor the Wolbachia symbionts. Grant ID: MCB-0604177

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该设备由MBL的当地科学家以服务能力使用。个人简短的研究描述如下： Sogin：我们的重点是与中心体相关的RNA及其可能的作用 在中心体的形成和功能中。 定量核酸和蛋白质中蛋白质的能力 生化制剂对于我们的研究至关重要。纳米体 使用微量的样本提供此功能，并 高灵敏度。 我们的研究目前由NIH GM075163资助 （克隆和分析中心体相关的RNA）。 Huber：地壳生物圈的采样不足，我们对存在于微生物的存在以及它们在这种动态的地球化学环境中如何分布的知识是碎片的。我们将使用来自Bio-RAD的DNA Engine Opticon实时PCR检测系统来量化来自整个太平洋（最终是大西洋）海洋中层状样品中的微生物基团。 我们没有资源在湾保罗中心进行定量PCR。 Bordenstein和Wernergreen实验室的成员成功地使用了Biocurrents机器，我们将在自己的协议开发中使用他们的专业知识。 Huber博士的L'Oreal奖学金将为MBL主管Gary Borisy博士提供的科学和初创企业计划的L'Oreal奖学金提供资金。 如果使用额外的赠款，我们会告诉您。 Wernergreen：这项研究探讨了细菌生活方式（自由生活与宿主相关）对基因组进化和种群遗传过程的速度和模式的影响。 该项目包括获取新的基因组序列数据，并为某些内共生物种的人群级变异进行测量。 我们依靠纳米体来确定分子工作中多个步骤中DNA的质量和浓度。 授予ID：2R01GM062626-06A1 Wernergreen：该项目整合了生态，生理和功能基因组方法，以检查强制性细菌互助中可塑性的机制。我们的核心目标是了解蚂蚁的内共生体如何应对宿主水平的变化。 我们使用QPCR机器来量化不同发育阶段和营养状况的宿主中细菌密度和基因表达模式。 Bordenstein：在过去的一年中，我们使用了纳米旋风来量化Wolbachia pipientis的DNA和RNA浓度，Wolbachia pipientis是无脊椎动物的细菌共生体，这些细菌也会引起象征性的丝状线虫感染的人类和河流百叶窗，这些丝虫具有丝状线虫感染的人类，这些丝虫具有沃尔巴赫菌的象征。 赠款ID：MCB-0604177