NANODROP AND PCR USAGE

这个子项目是众多研究子项目之一

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