NANODROP AND PCR USAGE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 该设备由MBL的当地科学家在服务能力中使用。个别简短的研究描述如下： Sogin：我们的重点是中心体相关的RNA及其可能的作用 在中心体的形成和功能中起作用。 我们对核酸和蛋白质进行量化的能力 生化制剂对我们的研究是必不可少的。纳米水滴 使用极少量的样本和 敏感度高。我们的研究目前由美国国立卫生研究院GM075163资助 (中心体相关RNA的克隆和分析)。 胡贝尔：地壳生物圈的样本仍然不足，我们对这种动态的地球化学环境中存在哪些微生物以及它们是如何在时间和空间上分布的知识是零散的。我们将使用Bio-Rad的DNA Engine Opticon实时PCR检测系统来量化整个太平洋(最终是大西洋)海底样本中的微生物群。 我们没有资源在贝保罗中心进行定量聚合酶链式反应。Bordenstein和Wernergreen实验室的成员已经成功地使用了BioCurrents机器，我们将在我们自己的协议开发中使用他们的专业知识。 这项技术的开发资金将由胡贝尔博士的L博士为科学中的女性提供的奥利弗奖学金和由MBL主任加里·鲍西博士提供的启动方案提供。如果使用额外的助学金，我们会让你知道。 Wernergreen：这项研究探索了细菌生活方式(自由生活与寄主相关)对基因组进化和种群遗传过程的速度和模式的影响。该项目包括获得新的基因组序列数据和调查选定内共生物种的种群水平变异。在我们的分子工作中，我们依靠纳米液滴在几个步骤中确定DNA的质量和浓度。 授权ID：2R01GM062626-06A1 Wernergreen：这个项目集成了生态、生理和功能基因组学方法，以检查专性细菌互惠共生中的可塑性机制。我们的中心目标是了解蚂蚁内共生菌如何在宿主水平上对变异做出反应。我们使用qPCR机来量化不同发育阶段和营养状况的宿主中的细菌密度和基因表达模式。 博登斯坦：在过去的一年里，我们使用纳米水滴来量化沃尔巴克氏菌的DNA和RNA浓度，沃尔巴克氏菌是一种无脊椎动物的细菌共生体，也会导致象皮病和河流盲症，人类感染了携带沃尔巴克氏菌共生体的丝虫线虫。 资助金编号：mcb-0604177

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