Provides equipment, methodology consultation, and reagents that assist in the introduction of transgenes for transient transfection, and in the localization of transgene and endogenous gene products. The most commonly used services are listed below, but, depending on investigator interest, other methods also can be offered as collaborative services.
Basic molecular biology and histology techniques
Techniques include, but are not limited to, animal perfusions, long-term preservation of tissue specimens, brain dissections, nucleic acids isolation and quantification, gel electrophoresis, Northern and Southern Blots, cellular fractionation, PCR and Western Blots. While these are routine methods in most laboratories, core equipment and experience help investigators devise, perform, and validate assays.
Expertise is provided for thionin/Nissl staining, widely used for revealing cell bodies and cytoarchitecture of brain areas, and for acetylcholinesterase staining, used to identify cholinergic neurons and fibers. Assistance with Hematoxylin and Eosin (cells, cytoarchiatecture), FluoroJade (degenerating cells), and Luxol Fast blue (myelin) staining also are available.
Antibodies are used to detect the presence of specific molecules (antigens) in tissue sections and provide the unique distribution pattern for a given antigen. IHC is performed both on slide-mounted sections or/and on fixed, free-floating sections, which also can be generated by the Core’s equipment. Core personnel have expertise with staining for >50 proteins, including, but not limited to transcription factors, neurotransmitter synthesis pathways, receptors, glial markers, cell adhesion proteins, inflammatory markers, cell signaling components, and synaptic proteins.
qPCR primer design and validation
Thoughtful primer design and validation are essential to ensure a sensitive and specific assay. The Core assists investigators in developing optimal qPCR primer sets for their needs. Once the assay is optimized, the investigator will perform the qPCR experiments in the Vanderbilt Technologies for Advanced Genomics Core (VANTAGE). vantage.vanderbilt.edu
These are usually applied to specific problems, where the investigator’s laboratory already has basic expertise in a technology but additional expert guidance is needed with some aspect. Core personnel also help with experimental design to maximize the likeliness of a successful outcome. Consultations at a later phase can evolve into collaborative services.