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ICMB Core Research Facilities
The ICMB core facilities support cellular and molecular biology research atThe University of Texas at Austin. The facilities offer a full range of services in nucleic acid and protein sequencing, peptide synthesis, mass spectrometry, protein purification and analysis, DNA microarrays, x-ray chrystallography, and transgenic - knockout mice. The core facilities are open on an equal basis to all faculty, regardless of whether they are ICMB members. The philosophy has been to keep the core facilities as comprehensive and accessible as possible to increase faculty, staff, and student research productivity.
DNA & Genomics Facility
The DNA Sequencing Facility provides DNA sequencing, fragment analysis, quantitative real-time PCR, nano-drop spectrophotometer, phosphor and fluor imaging, plate reader and automated liquid handling. Automated DNA sequencing is performed using capillary-based Applied Biosystems 3730 and 3130 DNA Analyzers. These instruments offer high throughput and sensitivity with a capability of handling more than 800 samples per day with reads >700 base pairs and a success rate of over 90%. The AB 3730 and 3130 are also used for the analysis of microsatellites, AFLP, SNP’s or any other fragment applications. Quantitative Real-Time PCR is run on an Applied Biosystems 7900HT. This instrument allows researchers to analyze gene expression, allelic discrimination and SNP analysis in 96 or 384 wells. The quantification of DNA, RNA and proteins using only 1 or 2 micro liters without a cuvette is performed on the NanoDrop spectrophotometer. The Typhoon Trio and Bio-Rad Molecular Imager FX measure and image radioactive signals from gels or membranes, and fluorescence from gels, membranes, TLC plates, or microtiter plates. Other instruments included in the DNA Facility are an Agilent Bioanalyzer, Beckman Biomek NX and FX pipetting robots, and a Beckman plate washer. A Berthold NightOwl was also added this year for low light imaging of luminescence or fluorescence in plants or animals. More information on services is available at http://www.icmb.utexas.edu/core/DNA/.
Microscopy & Imaging Facility
The ICMB Microscopy and Imaging Facility provides extensive microscopic equipment and services for ultra-structural analysis. The facility offers assisted use and training on its instrumentation and consultations on microscopy-and spectroscopy-related research. Equipment in the facility includes: scanning and transmission electron microscopes, confocal and wide-field fluorescence microscopes, cryo, paraffin, ultra microtomes, laser micro-dissection and stereology systems. The facility also provides state of the art image processing and analysis software. The Microscopy and Imaging Facility also manages the Flow Cytometry Laboratory which houses both a fluorescence cell analyzer and cell sorter. In collaboration with the IGERT Resource Center, we are able to provide access to additional equipment including ultra-violet, visible and infrared spectrometer, deconvolution and structured illumination microscopes. More information on services is available at http://www.icmb.utexas.edu/core/Microscopy.
Protein Microanalysis Facility
The ICMB Protein Microanalysis Facility provides research support for ICMB as well as other departments at UT. N-terminal protein/peptide sequencing, peptide synthesis and gel electrophoresis services are provided at accessable rates. Also available are HPLC, LC, capillary electrophoresis, mass spectrometry and analytical centrifugation systems. Researchers can sign up to use these instruments after a training session with the facility staff. The staff can also assist users in the preparation of samples for amino acid analysis performed off site. A new service, in conjunction with the Mass Spectrometry Facility, is the running of gels and in-gel digestions for protein identification. To limit keratin contamination, which can obscure MS data, both the gel and the digests are performed under clean conditions using a laminar flow hood. A two-dimensional gel electrophoresis service is currently in development. The instrumentation available in the facility includes an Edman-chemistry, N-terminal protein/peptide sequencer (ABI, Procise 492 cLc), peptide synthesizer (Rainin, Quartet), HPLC system (Beckman, System Gold), LC system (Amersham Pharmacia, AKTA), luminometer (Berthold Technologies, Mithras 940), capillary electrophoresis system (Beckman, PACE 5000) and an analytical centrifuge (Beckman, XL-I Protein Characterization System). More information on services is available at http://www.icmb.utexas.edu/core/Protein/.
Mass Spectrometry-Analytical Facility
The Mass Spectrometry Analytical Facility is administered jointly by ICMB, the College of Pharmacy and the UT M.D. Anderson Cancer Center Science Park. The core provides a variety of biomolecular analyses using sophisticated HPLC and mass spectrometry instrumentation. A state-of-the-art matrix assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF/TOF) is used for automated protein identification, with a sensitivity of <100 ng for high quality samples. The associated nano-LC spotting robot can detect peptides in the 100 fmol range. This instrument, as well as a MALDI-TOF, and electrospray-ion trap mass spectrometer with 1D and 2D-HPLC systems, is used for research into protein modifications and analysis of protein complexes in a variety of proteomics projects. Recent implementation of phosphopeptide enrichment protocols has enabled identification of novel sites of phosphorylation, and many other modifications have been characterized, including acetylation, methylation, inhibitor binding sites and chemical adduct formation. Analysis of metabolites, drugs, natural products and chemicals of interest is accomplished on an analytical HPLC coupled to an electrochemical detector or a gas chromatograph mass spectrometer (GC-MS). These instruments allow for sensitive detection of a variety of metabolites including 8-oxo-dG, a marker for oxidative DNA damage, neurotransmitter quantitation, and measurement on a variety of fatty acids. In order to sensitively detect multiple metabolites in a single sample, a triple quadrupole-ion trap (QTRAP) mass spectrometer has been added to the core. The TOF/TOF is located in MBB 1.420 while the other instruments are located in Pharmacy (PHR) 1.110. More information on services is available at http://www.utexas.edu/pharmacy/divisions/pharmtox/core/.
Mouse Genetic Engineering Facility
The Mouse Genetic Engineering Facility is located in the Animal Resource Center (ARC) and provides many services to the UT research community. The primary service is the production of genetically altered mice. This includes (1) the generation of transgenic mice (pronuclear injections), (2) gene targeting in mouse embryonic stem cells by homologous recombination (electroporation and selection), and (3) the subsequent generation of knock-out/knock-in mice (blasotcyst injections). Other services include embryo cryopreservation, long-term storage, and the recovery of frozen embryos, conversion of mouse strains to a pathogen free status, isolation of new embryonic stem cell lines from specific existing mouse strains, and assistance with timed-mating experiments for developmental studies. Our staff has expertise in obtaining blood samples from mice as well as tail vein injections and can assist investigators in this regard. The facility also maintains a specific pathogen free barrier for the housing and husbandry of mice. In early 2007, the generation of transgenic rats will be added to our menu of services. More information on services is available at http://www.icmb.utexas.edu/facilities/mouse/.
UT Microarray Facility
The goal of the UT Microarray Core Facilitiy is to help researchers interested in developing a custom microarray using a set of cDNA clones, genomic samples or oligonucleotides. Our primary aim is to provide cost-effective and time-efficient access to microarray technology, as well as an environment that facilitates effective analysis and sharing of microarray data. The Center for Systems and Synthetic Biology (CSSB) oversees the UTMCF, and is dedicated to providing microarray services to UT investigators as well as to off-campus users. The UTMCF provides a variety of services, including user or custom printing, array scanning, and training on the instruments in the facility. More information on services is available at http://cssb.icmb.utexas.edu/UTMCF.index.php.
Macromolecular Crystallography Facility
The Macromolecular Crystallography Facility, located in Welch Hall, allows users to solve the three dimensional structures of crystalline macromolecules, using X-ray diffraction methods. Dozens of high resolution protein structures have been solved using these facilities. The capacities of this center are now being expanded into a modern core facility. Current equipment includes two X-ray generators, detectors and cryo-cooling devices. The facility will be expanded in the coming year to include three state of the art detection systems. The Rigaku MicroMax 007 generator will have both high resolution and high flux mirrors. This generator will allow data collection on crystalline cells up to 300 Å is size and the high flux mirrors provide some of the strongest radiation outside of synchrotron sources. Cry-cooling will be available for all three detectors; this greatly facilitates collection of high resolution data. Associated with the core is a Phoenix, dual head, liquid handling robot. It uses extremely small volumes, down to 50 nl, and is ideal for large scale crystallization experiments. The new crystallography facility will be staffed to carry out structural analysis on a service basis, or to train and assist interested users in the collection, processing, and interpretation of X-ray data. More information on services is available at http://www.icmb.utexas.edu/xray/.