NANODROP AND PCR USAGE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 该设备由MBL的当地科学家在服务中使用。个人简要研究说明如下： Sogin：我们的重点是中心体相关RNA及其可能的作用 中心体的形成和功能。 在我们的细胞中定量核酸和蛋白质的能力 生化制剂对我们的研究至关重要。纳米滴 使用微量样品提供这种能力， 高灵敏度。 我们的研究目前由NIH GM 075163资助 （克隆和分析中心体相关的RNA）。 Huber：地壳生物圈仍然采样不足，我们对微生物存在以及它们如何随时间和空间分布在动态地球化学环境中的知识是零碎的。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）海底样品中的微生物群。 我们没有资源在海湾保罗中心进行定量PCR。 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Huber博士的欧莱雅科学界妇女奖学金和MBL主任加里Borisy博士提供的启动包提供。 如果使用额外的赠款，我们会让你知道。 沃纳格林：本研究探讨了细菌生活方式（自由生活与宿主相关）对基因组进化和群体遗传过程的速率和模式的影响。 该项目包括获得新的基因组序列数据和调查选定的内共生物种的种群水平变化。 我们依靠nanodrop在分子工作的几个步骤中确定DNA的质量和浓度。 授权ID：2 R 01 GM 062626 - 06 A1 沃纳格林：该项目整合了生态学、生理学和功能基因组学方法，以研究专性细菌互利共生中的可塑性机制。我们的中心目标是了解蚂蚁的内共生体如何在宿主水平上对变化做出反应。 我们使用qPCR机器来量化不同发育阶段和营养状态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Nanodrop来量化Wolbachia pipientis的DNA和RNA浓度，Wolbachia pipientis是一种无脊椎动物的细菌共生体，也会导致感染了携带Wolbachia共生体的丝虫的人类的象皮病和河盲症。 授权ID：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