ICMB Core Research Facilities

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 and Metabolite Analysis Facility

The Protein and Metabolite Analysis Facility provides a variety of biomolecular analyses using sophisticated mass spectrometry instrumentation, as well as other protein services and instrumentation. It is administered jointly by ICMB, the College of Pharmacy and UT M.D. Anderson Cancer Center-Science Park. The newest addition is Luminex assays using multiplexed antibody-bead technology to provide sensitive quantitation of cytokines, hormones, and other analytes in a 96 well format. A MALDI-TOF/TOF mass spectrometer with a nano-LC spotting robot is used for identification and quantitation of proteins. Proteolytic digest is available as a core service. Protein modification studies are conducted in collaboration with research labs. The phosphopeptide enrichment protocol enables identification of novel sites of phosphorylation, and many other modifications have been characterized, including acetylation, methylation, inhibitor binding sites and cysteine oxidation. An LCQ provides protein/peptide molecular weight determination. Quantitative analysis of metabolites, drugs, nutrients, and natural products is conducted on QTRAP, GC-MS, and HPLC-EC instruments. Multiple metabolites are monitored in a single run, with methods in place for prostaglandins and fatty acid methyl esters. A MALDI-TOF is available for self-service work. N-terminal protein/peptide sequencing, peptide synthesis and gel electrophoresis services are provided on site. Also available are HPLC, protein purification chromatography, crystallization robot, microplate reader, lyopholizer and analytical centrifugation. Researchers can sign up to use these instruments after a training session with the facility staff. 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.

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/core/xray/.

Genomic Sequencing and Analysis Facility

The Genomic Sequencing and Analysis Facility at the University of Texas at Austin is located in room 4.102 of the MBB building, which houses the ICMB. We perform next-gen sequencing and microarray studies for UT researchers from any department and from external research entities, both public and private. We are supported by UT's Texas Institute for Drug & Diagnostic Development and the Center for Systems and Synthetic Biology. More information on services is available at https://wikis.utexas.edu/display/GSAF/.