The purpose of this Core is to provide molecular analysis services to the investigators in this Program Project. Western immunoblot; Northern and in situ hybridization; and quantitative, competitive reverse transcriptase polymerase chain reaction (RT-PCR) analyses; real-time PCR; microarray; DNA fragmentation analysis; electromobility shift assay; enzyme assays for acetylcholine (AChE)-related enzymes and for ICE; and cytokine bioactivity analysis of cytokine cycle proteins are available. In addition, analysis of other proteins and their encoding mRNAs that may be of interest (e.g., other cytokines, growth factors, or structural proteins) will be available upon request. S100beta and GFAP ELISAs are available, and other ELISAs can be constructed, for rapid screening of large numbers of samples. As a rapid screen for specific mRNAs in cell cultures, micro-in situ hybridization is also available. The tissue to be analyzed will be from human and rodent brain and muscle (Projects by Griffin, Gentleman, and Peterson) and from cell cultures (Projects by Barger and Peterson). Data from analyses performed in this Core will be provided to the Project investigators as both raw data and preliminary analyzed data. For example, data from Northern and Western immunoblot analyses and RT-PCR will be returned to the investigator as raw data (pictures of gels and films of transfers) and as ng/mug or ng/mul of extract, relative to total protein, total RNA, G3PDH, or ribosomal RNA that can be hybridized. For real time PCR, cycle-by-cycle detection of PCR product accumulation gives direct quantitative results with no additional analysis steps required. Data for microarray analyses will be provided as relative fluorescence intensity following hybridization. Data from ELISAs will be returned as raw data in the form of optical density units per sample, together with a standard curve generated at the same time, and calculation of ng/mug protein and ng/mul extract. Following in situ hybridization, sections or cell cultures will be returned to the investigator. Data from micro-in situ hybridization of cultures will be returned to the investigator as cpm/well relative to total polyadenylated mRNA. Verification of an values from services performed in this core will be archived in the Core's freestanding computer database.
Showing the most recent 10 out of 140 publications
