GW Biomarker Discovery and Analysis Core Facility
The GW Biomarker Discovery Facility provides resources for the detailed analysis of gene and protein expression in the nervous system and other services.
Quantitative PCR services
The BDAF has a Bio-Rad CFX-384 qPCR platform for analyzing transcript levels in a 384 well format. The 384 well platform allows for the simultaneous analysis of multiple transcripts in a single experiment, and reduces reagent costs as compared to a similar 96 well qPCR platform.
To increase the accuracy, repeatability, and throughput of qPCR assays, the BDAF has a Eppendorf EpMotion 5070 liquid handling system, to facilitate the setup of qPCR experiments.
The accuracy and throughput of this qPCR system will facilitate many aspects of biomarker analysis, from the precise measurement of expression levels for one or two genes from many tissue samples, or for validating expression changes detected in a microarray experiment.
The BDAF can provide cDNA samples from a library of mouse tissues (embryonic and postnatal brain; postnatal organ tissues); in addition, we can work with researchers to produce high-quality cDNA from tissues.
In situ hybridization
The BDAF provides in situ hybridization analysis services on cryostat sections, for localization of expressed transcripts in developing and adult tissues. Expression analysis by ISH allows for the localization of expressed genes, to identify biomarkers that are associated with specific cell types or tissues, and can also be used to assess changes in a biomarker corresponding to a disease state or genetic lesion.
BDAF staff will work with investigators to amplify, clone, and subclone appropriate The BDF also has a library of probes available, with emphasis on genes important for mouse neural development and patterning.
The BDAF can provide ISH-ready slides from a tissue-bank of standard mouse tissues (E14 embryo, Adult brain); or we can provide assistance in producing cryostat sections of tissue suitable for ISH.
The BDAF has a Licor Odyssey infrared imaging system that is capable of high-sensitivity two-channel fluorescent scanning of western blots and gels. Under optimal conditions, the scanning system is capable of producing quantitative measurements of protein levels for multiple proteins (from 2 to 4 or more individual proteins). The availability of two independent fluorescent channels, and the use of direct fluorescent detection allows for accurate quantitative comparisons of protein levels from tissue samples.
Luminescence and Fluorescence microplate assays
The BDAF has a Thermo Varioskan microplate reader, which is capable of highly sensitive and accurate assays of luminescence and fluorescence. The Varioskan is monochrometer based, so it is adaptable for different fluorescence assays simply by setting the desired wavelength.
Primary culture and electroporation laboratory (Ross Hall 649)
Complete facilities, including two dissecting stations in laminar flow air curtain hoods, two CO2/temperature controlled incubators, Leica dissecting fluorescence microscope, Leica vibratome for live slice preparation, picospritzer for specimen injection, and square wave electroporator are available for use.
Cell culture and ES cell laboratory (Ross Hall 648)
Complete facilities, including two sterile tissue culture hoods, centrifuges, two CO2/ temperature controlled incubators, stocks of standard cell lines (HEK, L, MDCK), Mouse ES cells (both S129 and C57b6) for differentiation, preparative centrifuges, and other standard cell culture equipment are available for use.
Other services and facilities:
- Rapid Polymorphism detection by High-Resolution Melt Curve Analysis
- Luciferase-based bioassays for developmental signals (Shh, Bmps, Retinoic Acid, etc.)
- Cell line development (Tetracyline regulated expression systems)