Iowa State University



Animal Gene Transfer FacilityLocation:1270 Kildee HallHours:8 a.m. to 5 p.m. weekdays, by appointmentContact: Jason Ross, Professor-in-Charge 515-294-8647, jwross@iastate.eduFax:515-294-4471The facility provides a variety of services to individuals who wish to utilize transgenic animals as part of their research program. The facility maintains equipment necessary for the creation of transgenic animals via microinjection or somatic cell nuclear transfer and is available to life science researchers for technical service and for training of a limited number of individuals. All work to be performed in this facility must be scheduled with the professor-in-charge, as the facility is not staffed full-time.Equipment housed in the facility includes two microinjection work stations, micropipette pullers, a microforge, stereo-zoom microscope and CO2 incubator. Additional supporting equipment located in nearby laboratories to which access may be granted includes a heated microcentrifuge, tri-gas incubators, stereo-zoom microscopes, inverted microscope, biosafety cabinet, osmometer, pH meter, embryo freezing machine, cell fusion machine, gel electrophoresis equipment and UV transilluminator.Atmospheric Air Quality LaboratoryLocation: 3115 National Swine Research and Information CenterHours: 8 a.m. to 5 p.m. weekdaysContact: 515-294-9174 FacilityJacek Koziel, Professor-in-Charge515-294-4206, koziel@iastate.eduFax:515-294-4250Web:abe.iastate.edu/odorThe Atmospheric Air Quality Laboratory is situated at the National Swine Research and Information Center and serves as a focal point for collaborative research and training in the area of air quality engineering and livestock odor. The laboratory specializes in development and testing of odor and gaseous emissions mitigation technology, continuous air quality monitoring, environmental analysis, quantification of organic compounds with gas chromatography and mass spectrometry, and simultaneous chemical and olfactometry analyses.Livestock odor and gaseous emissions mitigationThe lab offers lab-scale and pilot-scale development and testing of odor and gaseous emissions mitigation based on physical, chemical and biological approaches. Testing of the treatment efficacy targeting odor, ammonia, hydrogen sulfide, volatile organic compounds, greenhouse effect gases (CO2, CH4, N2O) are conducted. Full scale tests are conducted at university and commercial farms.Air quality monitoring for odor, gases, particulate matter and greenhouse gases at livestock operationsThe laboratory is equipped with a variety of real-time analyzers for gaseous ammonia, hydrogen sulfide, carbon dioxide, methane, nitrous oxide, volatile organic compounds, particulate matter and ozone. It is also equipped to conduct field air sampling and analysis of organic gases with sorbent tubes and SPME.Wine flavor, aroma and nutraceuticalsThe lab specializes in state-of-the art chemical and sensory analysis of wine flavors, aromas and nutraceuticals. Chemical method detection limits are between part-per-billion and part-per-quadrillion levels.Multidimensional GC-MS-olfactometryThe unique MDGC-MS-O system includes a custom-made GC-MS system with heat-cutting capability based on a Dean switch capable of simultaneous chemical and olfactometry analyses.Solid phase microextraction (SPME)SPME is used for extractions of gases emitted from a variety of samples, including livestock and poultry manure, air, food, beverage, packaging, bio-based products, live insects, plants, breath and general applications related to quality control and quality assurance.Non-destructive in vivo sampling and analysis of volatilesThe lab is equipped for technologies for non-destructive in vivo sampling and analysis of volatiles from insects, plants, fruits, and breath.Biofuels analysesSampling and analysis of volatiles and semivolatiles at elevated temperatures from various processes associated with biofuels production and utilization are available.Atomic Force Microscopy FacilityLocation:0122 Molecular Biology BuildingHours: 8 a.m. to 5 p.m., weekdays; 24-hour access after complete trainingContact:Curtis Mosher, Manager515-294-6513, cmosher@iastate.eduFax: 515-294-7134Three atomic force microscopes are available for use in the Roy J. Carver Laboratory for Ultrahigh Resolution Biological Microscopy of the Institute for Combinatorial Discovery.Digital Instruments DimensionTM 3000 Scanning Probe MicroscopeThe Dimension 3000 scanning probe microscope (SPM) brings all SPM techniques into a single platform and handles a wide range of sample sizes and types. A rigid, low vibration construction of the Dimension 3000 SPM ensures the highest quality images and measurements.Samples up to eight inches in diameter can be scanned in ambient air or fluids using the Dimension 3000 SPM. The Dimension 3000 SPM requires little or no sample preparation, and the simple vacuum mounting system allows easy and convenient setup. Superior linearity and resolution in all three dimensions are obtained, even for large samples. Integrated top-view video optics with motorized zoom and 1.5 ?m optical resolution help identify areas of interest for detailed scanning quickly and easily. Changing scanning techniques, for example from AFM to STM, requires no tools.The NanoScope IIIa system controller is a main part of the SPM system, providing the software and electronics that drive the microscope. Digital tracking and feedback control insure accuracy and speed at all scan sizes and positions on the sample.Digital Instruments MultiMode with Tapping Mode TM The MultiMode system features multiple scanners that permit the user to tailor the system for individual research. Scanners with large scan ranges up to 120 microns on the X–Y axes and a Z range up to 6 microns, as well as high-resolution scanners with 0.5 microns X–Y axes and submicron Z range are available. The vertical-engage “JV” and “EV” scanners allow the tip to be positioned at any point on the surface, without adjusting for lateral movement during approach. The MultiMode is controlled with a NanoScope IIIa controller. This controller provides 16-bit resolution on all three axes, with three independent 16-bit digital-to-analog converters (DACs) in X and Y for control of the scan pattern, scaling and offset. This configuration provides 16-bit resolution of the lateral scanning motion at any scan size.Digital Instruments DimensionTM 3100 Scanning Probe MicroscopeThe Dimension 3100 is controlled with a Nanoscope IV controller. The NanoScope IV features up to ten-times-faster scanning, as well as increased functionality, bandwidth, flexibility and expandibility. Biomolecular Nuclear Magnetic Resonance FacilityLocation:0208 Molecular Biology BuildingHours: 9 a.m. to 5 p.m. weekdays or by appointmentContact: Bruce Fulton, Manager515-294-2139, bfulton@iastate.eduWeb: public.iastate.edu~bfulton/nmr_home.htmlThe Nuclear Magnetic Resonance (NMR) Facility is supported by the Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology. The facility currently operates Bruker Avance 700 and 500 spectrometers, both capable of performing a broad range of modern multi-nuclear, multi-dimensional NMR experiments on biomolecules. The 700 is equipped with an H/C/N cryoprobe, an H/C/BB conventional probe and an H/BBX/BBY-MAS solids probe. The 500 is equipped with an H/C/N cryoprobe as well as H/C/N and BB/H conventional probes.The facility has computational resources for processing and analyzing NMR data and obtaining molecular structures. The facility provides consultation on the application of NMR to solve research problems. NMR data can be acquired and interpreted as an analytical service. Projects larger in scope can be pursued on a collaborative basis. In the latter case, the facility will provide training and guidance for researchers to operate instruments and interpret data.Center for Crops Utilization ResearchLocation: 1041 Food Sciences Building (Administration)1955 Food Sciences Building (Pilot Plant Office)1327 U Avenue, Boone, IA (BioCentury Research Farm)Hours:8 a.m. to 5 p.m. weekdays (fall, winter, spring)7:30 a.m. to 4:30 p.m. weekdays (summer)other hours by arrangementContacts:Lawrence Johnson, Director, 515-294-0160, ljohnson@iastate.eduHui Wang, Pilot Plant Manager, 515-294-3572,huiwang@iastate.eduAndrew Suby, BioCentury Research Farm Manager515-294-6300, asuby@iastate.eduFax:515-294-6261Web:ur.iastate.edu, biocenturyresearchfarm.iastate.eduThe Center for Crops Utilization Research (CCUR) is a resource to assist ISU researchers and external businesses in developing new value-added processes, products and markets for Midwest crops, especially corn and soybeans. CCUR strives to add value to grain and other crop-derived materials by conducting grant and contract research; offering short courses, workshops, seminars and training experiences; providing analytical, pilot plant processing, and consumer evaluation services; providing technical consulting services and information retrieval; and operating small-business incubator services. The center has grain, food and material processing equipment in state-of-the-art laboratory and pilot plant facilities.The pilot plant facilities include a 5,000 square-foot wet-processing pilot plant (for soy protein isolation, corn wet milling, brewing, etc.); a 2,600-square-foot dry-processing pilot plant (for dry corn milling, drying, grinding, sieving, etc.); a 900-square-foot hazardous solvents extraction facility (vegetable oil extraction from grain, plant material extractions); a 3,000-square-foot product development laboratory (for plastic extrusion, molding, and film blowing; building materials processing; etc.); and various process development and analysis laboratories (for chromatography, grain analysis, vegetable oil refining, baking, laboratory-scale process development, etc.). A small theater and conference facilities are available for technology transfer UR partly administers and works closely with the ISU Fermentation Facility, the Iowa Grain Quality Initiative and the Grain Quality UR also manages the BioCentury Research Farm (BCRF), the first integrated biomass-to-biofuel production and processing facility in the world, located seven miles west of the ISU campus on US Highway 30. It offers opportunities for pre-commercial-scale research in biomass feedstock production, harvest, transport, storage, preparation, biorefinery processing and laboratory testing. Field plots for crop production trials, field equipment modification facilities, biomass storage facilities and a prototype biorefinery conversion facility are available for on- and off-campus users. The biomass processing facility houses three, 2,700-square-foot pilot plant processing trains which include biomass chemical fractionation and other technologies, four laboratories totaling 2,100 square feet, and 1,000 square feet of office space. The BCRF also has 4,200 square feet of dry biomass feedstock storage and 420 square feet of cold storage. Services include biomass feedstock preparation (sizing and drying) and fermentation production of fuels and industrial chemicals (50, 200, 500 and 1,000 liter capacities).Chemical Instrumentation FacilityLocation: Suite 1234 Hach HallHours:24 hours a day, seven days a week; professional help available from 8 a.m. to 5 p.m. weekdaysContacts:Steve Veysey, Manager, sveysey@iastate.edu515-294-5805NMR Lab: Sarah Cady, Shu Xu, 515-294-5975Mass Spec Lab: Kamel Harrata, 515-294-5544 X-Ray Lab: Arkady Ellern, 515-294-6419Web:cif.iastate.eduThe Chemical Instrumentation Facility has more than five million dollars worth of analytical instrumentation available to faculty, graduate students, industry and other educational institutions. The staff of five highly-qualified professionals supports university research by keeping the analytical equipment available and operable and by providing application support and user training. Services available to the research community include the following:Magnetic ResonanceSix NMR spectrometers of varying frequencies from 300 to 600 MHz are available. Magnetic resonance spectroscopy allows the use of atomic nuclei as magnetic probes within a molecule. Chemical analysis and spatial orientation are determined by using this technique. A fully-equipped EPR system also is available. Equipment located in the facility includes a Bruker Avance II 600 (solids) NMR, a Bruker Avance III 600 (solution) NMR, a Bruker DRX-400 NMR, an Agilent MR-400 NMR, a Varian VXR-300 NMR, a Varian VXR-400 NMR and a fully-equipped Bruker E-580 FT-EPR.Mass SpectrometryThe mass spectrometry lab is equipped to provide both low- and high-resolution GC-MS on mixtures and high-resolution measurements for determining the elemental composition of pure samples and multi-component mixtures. Electrospray and APCI are used routinely for the ionization of medium and higher molecular weight compounds, including synthetic organics and compounds of biological origin. Three dedicated LC-MS instruments are available for routine work and special projects. MS-MS (parent-daughter relationship) experiments are routine. Equipment located in the facility includes a Finnigan Magnum ITD GC-MS, a Waters Micromass GCT-MS, a Shimadzu LCMS2010, a Finnegan LCQ LCMS, and an Agilent 6540 Q-TOF LC MSMS.Preparative HPLCThe Agilent PrepStar SD-1 preparative HPLC system is capable of flows ranging from 1-2 mls per minute (analytical) to 200 mls per minute (preparative). The system includes two pumps, a dual-wavelength detector, fraction collector and complete data system for instrument control and data processing. Both semi-prep (9 mm i.d.) and preparative (21 mm i.d.) columns are available for use for an additional fee.X-ray DiffractionA completely-equipped X-ray diffraction laboratory provides instrumentation for the study of the molecular structures of small molecules and powders. The equipment includes Bruker APEX II CCD and SMART 1000 single-crystal diffractometers equipped with low-temperature devices and Scintag XDS-2000 and Rigaku Ultima 4 powder diffractometers available for general use. The facility’s crystallographer provides complete reports that are suitable for publication.SpectrophotometryA variety of spectrophotometers is available for routine use in the facility. These instruments provide fingerprint spectra for characterizing and identifying compounds. These instruments currently include a Bruker IFS 66V FT-IR, a Hewlett-Packard HP-8453 Diode Array UV-Vis, a Cary 100 Bio double beam UV-Vis, and a Jasco J-710 circular dichroism spectrophotometer.Isothermal Titration Calorimetry (ITC)A Microcal ITC is available to study the thermodynamics of chemical reactions initiated by the addition of a binding component. It is often used to characterize biomolecular interactions.Elemental AnalysisA Perkin-Elmer Model 2400 Series II CHN/S elemental analyzer is available for sample submission or for investigator use. Normally, the instrument is configured for carbon, hydrogen and nitrogen, but sulfur also can be analyzed upon special putationIn addition to computer systems associated with the instrumentation, numerous PCs and workstations are available for network-based data processing and parative Pathology Core ServiceLocation:2720 Veterinary MedicineHours:By appointment, Monday through Friday 8 a.m. to 5 p.m.Contact:Jesse M. Hostetter, 515-294-0953jesseh@iastate.edu The Comparative Pathology Core Service (CPC) provides pathology services and consultation for investigators working with animal models of human and animal diseases. The veterinary pathologists in the CPC provide services for all phases of research involving animal models. This includes animal model selection, gross and microscopic tissue examination, clinical pathology evaluation and interpretation and support for grant applications. Consultations with a CPC pathologist prior to project initiation will often provide the most effective pathology support.In addition, the core can develop and implement ancillary techniques which include immunohistochemical methods, fluorescence microscopy, laser capture microdissection and photomicroscopy. CPC costs are based on histopathology lab services and pathologists’ effort, which will vary based on the needs of the project. More detailed information can be provided by contacting the CPC.Confocal Microscopy and Multiphoton FacilityLocation:0071 Roy J. Carver Co-Laboratory0117 Molecular Biology BuildingHours:8 a.m. to 5 p.m. weekdays and as arranged 24-hour access after complete trainingContacts:Margie Carter, Manager, 515-294-1011mcarter@iastate.edu Fax: 515-294-7629 The Confocal Microscopy and Multiphoton Facility of the Office of Biotechnology and the Plant Sciences Institute have confocal and multiphoton microscopes available for use by internal and external researchers.The confocal microscopes are located in 0117 Molecular Biology Building and 0071 Roy J. Carver Co-Laboratory. Researchers can choose which microscope best fits their research needs.Confocal microscopes remove out-of-focus fluorescent light from the image, allowing clearer imaging of the sample, including the ability to view structures and components that were obscured by excess fluorescence that would be generated by standard fluorescence microscopes. Some applications of confocal microscopy include fluorescence microscopy when spatial distribution of cellular or other structures is important to the research being conducted, acquisition of a stack of images that can be used for 3D-reconstruction, viewing structures in thick tissue and removing background and cross-talk fluorescence or fluorescence from other structures that obscure viewing of the desired structure.Molecular Biology BuildingThe confocal microscope in the Molecular Biology Building allows for real-time optical sectioning of fixed and living specimens, providing significant improvements in optical contrast and resolution over traditional light and fluorescence microscopy. The facility is equipped with a Leica SP5 X MP confocal/multiphoton microscope system with an inverted microscope front end. New technology available on this system includes a white light laser, IR laser and optical parametric oscillator (OPO). Researchers are no longer limited to three or four lasers at three or four fixed wavelengths but can tune the white light laser to any wavelength between 470 and 670 nm, which has a similar effect to having 200 lasers and 200 wavelengths. This allows researchers to select the excitation wavelength that is best suited to their samples and will result in maximum fluorescence emission. On the emission side, the system has an Acousto-Optical Beam Splitter (AOBS) which allows researchers to precisely set the emission wavelengths they would like to capture rather than be limited to preset emission ranges determined by fixed filter sets. Both of these technologies help to separate fluorescence signals with close excitation and/or emission spectra.Other features include faster scanning, higher resolution and increased sensitivity. Researchers will now be able to work with live cells over an extended period using live cell equipment, including a heated stage with microcontainment or macrocontainment system, active gas regulation, cell cultivation chamber and microinjection system. For capturing images of fast moving samples or events that occur in a fraction of a second, the resonant scanner enables video rate scanning with speeds up to 16,000 lines per second. In addition, the system has FRET, FRAP, timelapse, hyperspectral signal separation software, 3D imaging, colocalization, deconvolution, region of interest (ROI) scanning, brightfield and DIC capabilities. A 405 laser provides UV excitation, and an Argon laser provides CFP excitation and extra power for bleaching. The system also has an X-Y scanning stage and automatic composition software to capture high resolution images of samples that are too large to fit within one field of view and a color digital camera.The system allows users to easily switch between confocal and multiphoton mode. In multiphoton mode, users can image deeper into their samples than is possible with confocal systems. The higher excitation wavelengths are less damaging to live samples, as well. The OPO expands the excitation range of the multiphoton laser to include the red and high red ranges, enabling use of essential fluorophores such as mCherry, DsRed2, mRFP, Texas Red, Alexa Fluor 660 and others. Researchers are able to precisely set the emission wavelengths they would like to capture. DNA FacilityLocation:1184 Molecular Biology BuildingHours:7:30 a.m. to 5 p.m. weekdays or by appointmentContacts:Gary Polking, Manager515-294-1813, 515-294-7629 (fax)E-Mail:polking@iastate.edu(DNA Sequencing) dnaseq@iastate.edu (DNA Synthesis) isu_oligo@iastate.edu Web: dna.iastate.eduThe DNA Facility of the Office of Biotechnology performs Sanger DNA sequencing, next-gen (Illumina) sequencing, plant genomic and plasmid DNA extraction, DNA and RNA characterization, automated fluorescent genotyping, DNA synthesis and primer walking, quantitative and digital PCR, including Fluidigm BioMark HD genetic analysis, and GC analysis of oil seeds.Sanger Sequencing (dna.iastate.edu/sangersequencing.html)Sequencing samples are run on the DNA Facility’s Applied Biosystems 3730xl DNA Analyzer. The AB 3730xl provides up to 900 bases of usable sequence data per reaction and is capable of processing 12 sets of 96 samples in a 24-hour period. DNA can be sequenced as plasmid, lambda, cosmid or BAC DNA, or as PCR products, and custom primers can be used with all types of templates. Clients submit sequencing orders using the OnCore software. This software allows clients to track the progress of their orders and automatically notifies them when their data are ready to download. A four-color printout of the data is provided. When no problems are encountered with a template, the results are generally returned within 48 hours after receipt of the samples. The facility also provides a primer walking service (dna.iastate.edu/primerwalking.html).For clients who have high-throughput sequencing projects, samples can be submitted in 96-well format. The facility should be contacted prior to submission of samples in 96-well format.Next Gen Sequencing (dna.iastate.edu/nextgensequencing.html)The Iowa State DNA Facility operates both an Illumina HiSeq2500 and an Illumina MiSeq. These instruments can produce up to 180 million (HiSeq high output mode), 150 million (HiSeq rapid run mode) and 15 million (MiSeq) sequencing reads per lane, making them ideally suited for many applications, including ChIP-seq, genome sequencing, mRNA and small RNA sequencing, and amplicon sequencing. Clients can choose various read lengths and either single or paired-end sequencing. Depending on the application, up to 96 samples can be multiplexed per lane. The facility provides library construction services for all Illumina applications and also accepts client-prepared libraries.Genotyping (dna.iastate.edu/genotyping.html)The facility can process a number of marker types including SSRs, AFLPs, SNPs, ISSRs, and T-RFLPS using an Applied Biosystems 3730 DNA analyzer to electrophorese samples and collect the gel image. Each sample can have as many markers as the client can identify. The data are analyzed using the Applied Biosystems GeneMapper Analysis software, and electronic files are provided via the facility server.Oligonucleotide Synthesis(dna.iastate.edu/oligosynthesis.html)The DNA synthesis service synthesizes DNA oligomers in two scales– 50-nmol and 200-nmol– and can make modified oligomers such as the fluorescent primers used in genotyping applications. Oligos are synthesized using a BioAutomation MerMade-192 DNA synthesizer. In addition, primer design for primer walking sequencing projects is available (dna.iastate.edu/primerwalking.html).DNA Template Preparation and Miscellaneous ServicesPlasmid Preparation Service (dna.iastate.edu/plasmiddnaextraction.html) The facility offers two different 96-well format plasmid preps- Millipore’s Montage prep and a lower cost screening prep.Plant Genomic DNA Extraction Service (dna.iastate.edu/plantdnaextraction.html) The facility performs plant genomic DNA extraction using the AutoGen 740 Automatic DNA Isolation System, a fully automated instrument that is capable of extracting 192 samples per day. Extraction can be performed on either fresh, or preferably, lyophilized material from a variety of plant tissues.Plant Tissue and Seed Grinding Service (dna.iastate.edu/tissuegrinding.html) Plant tissues and seeds are ground in polycarbonate tubes using stainless steel ball bearings. Entire, intact seeds can be ground into fine powder.DNA and RNA Characterization(dna.iastate.edu/bioanalyzer.html)The facility has two instruments available to assist with nucleic acid sizing and quantification. The Agilent Bioanalyzer is best suited to smaller numbers of samples; the Advanced Analytical Fragment Analyzer can quickly and economically process larger numbers of samples. New users will receive initial instrument training.Quantitative and Digital PCR (dna.iastate.edu/quantitativepcr.html)The DNA Facility maintains two quantitative, real-time PCR instruments — the Applied Biosystems StepOnePlus and the Stratagene Mx3005 — and will accept jobs on a ready-to-run basis. Applications include gene expression studies, validation of microarray data, allelic discrimination, SNP analysis, and screening for GMOs. For clients with projects that require a higher throughput, the facility has available the Fluidigm? BioMark? HD system. This instrumention utilizes microfluidic technology to accurately combine reagents and samples to perform thousands of individual nanoliter-scale reactions in a single run. DNA staff are also available to advise or assist clients at any point in the experimental process from initial project design through chemistry and material selection and data analysis.GC Analysis of Oil Seeds (dna.iastate.edu/seedoilanalysis.html)The facility provides soybean fatty acid analysis as a service for researchers on and off campus. Relative concentrations are determined in percent of palmitic, stearic, oleic, linoleic, and linolenic fatty acids by measuring fatty acid methyl esters using the facility’s Hewlett Packard 5890 gas chromatographs.Doubled Haploid FacilityLocation: 1204 Agronomy HallHours:8 a.m. to 5 p.m. weekdaysContacts:Thomas Lübberstedt, Professor-in-Charge, 515-294-5356, thomasl@iastate.eduUrsula K. Frei, Research Scientist, 515-294-2756,ufrei@iastate.eduFax: 515-294-5506Web: Doubled Haploid Facility (DHF) provides haploid doubling for maize to ISU scientists, as well as off-campus academic researchers and breeders. The start-up phase is supported by the Department of Agronomy which provides use of its laboratories, greenhouses and nursery.The development of homozygous lines is an important, but time-consuming, process in plant breeding and research. The induction and subsequent doubling of haploids is an efficient alternative to generate homozygous offspring in two generations.The mission of DHF is to:provide expertise and service in the production of doubled haploid lines in maizeimprove the technology in order to get higher success rates and lower coststeach and train scientists and studentsThe production of doubled haploid lines consists of three main steps:haploid induction by pollination with inducer lines,selection for haploid kernels,doubling of haploid plants and subsequent seed productionhaploid inductionFor haploid induction, maternal inducer genotypes, such as RWS described by Roeber et al. 2005 are used. In the experiments, the average induction rate (IR) is about 8% in diverse plant materials.Haploid Kernel SelectionKernels with a haploid or F1 embryo can be distinguished by means of the expression of the dominant anthocyanin marker gene R1-nj. The expression of this gene provides an anthocyan pigmentation of the embryo and the endosperm. Kernels with a pigmented endosperm and a nonpigmented embryo are selected as haploids.DoublingSpontanous chromosome doubling occurs but is dependent on the genotype. To obtain more consistent doubling rates, plants are treated with colchicine following the procedure (method II) described by Eder et al. 2002. Doubling rates (DR) of up to 20% are realized. On average, 8.4% of the initially grown haploid seeds resulted in a doubled haploid line (DHL) in 25 diverse donor populations from 10 different breeding companies.The production of the double haploid lines requires two generations. For one generation, a winter nursery in Chile is used.Environmental Engineering Research LaboratoryLocation: 123 Town EngineeringHours:8 a.m. to 5 p.m. weekdaysContacts:Facility 515-294-8579, 515-294-8216 (fax)Shiwu Sung, Professor-in-Charge515-294-3896, sung@iastate.eduDavid G. Schoeller, Assistant Chemist515-294-8579, dgschoel@iastate.eduThe laboratory is a service of the Civil, Construction and Environmental Engineering Department and provides chemical analysis and related training and consultation services in support of university-sponsored research. Documented quality control receives top priority and is made available to researchers wishing to verify the quality of results.Facility staff can function as consultants to assist the ISU research community by preparing quality assurance plans for research proposals, configuring computer systems for data acquisition and manipulation, training departmental personnel in analytical instrumentation and methodology, implementing chemical hygiene plans and preparing specifications for instrument purchases. Most of the laboratory’s major instrument systems are available for use by researchers who wish to do their own analytical work. The laboratory provides training and supervision for those researchers.Atomic Absorption SpectrophotometryFlame atomic absorption and emission are available. A 150-position autosampler and a GBC 932 Plus Atomic Absorption Spectrophotometer can be used to determine metals in large numbers of samples.Automated AnalysisA Seal Analytical AQ2+ automated discrete analyzer is available for unattended automated analysis of nitrogen, phosphorus, silica, sulfate, chloride and cyanide.A BD-40 block digester is available for simultaneous, semi-automated Kjeldahl nitrogen and total phosphorus digestions. Data collection, computation and reporting are carried out with a microcomputer data station.Mercury AnalysisA Leeman Labs Hydra AF Gold Plus automated mercury analyzer is available to analyze for trace levels of mercury. The analyzer uses cold vapor atomic fluorescence with dual stage gold amalgamation and dual detectors.GC/MSA Varian Saturn 2100 ion trap GC/MS system that includes MS/MS, SIS, and CI capabilities is available. The system includes an autosampler and GC/MS data station.Gas ChromatographyFour gas chromatographs served by EZ Chrom Elite Chromatography Data Systems provide a wide range of options for organic analysis. FID, ECD, TCD, ELCD, PID and NPD detectors are available. Autosamplers are available for liquid injection, as well as purge-and-trap sampling. Chromatographs are equipped for operation with packed, capillary or Megabore columns. Sample preparation techniques include continuous or discrete liquid-liquid extraction for water samples and sonication or Soxhlet extraction for soil, tissue and other solid samples.Carbon AnalysisA Shimadzu TOC-Vws TOC Analyzer is available to analyze carbon in liquid samples by the UV-promoted persulfate oxidation method.OtherThe laboratory is equipped for most types of wet chemical analysis, including related spectrophotometric and potentiometric methods. Membrane filter techniques are used for bacterial testing.Fermentation FacilityLocation:1621 Food Sciences Building (main location)2058 Sweeney Hall (satellite location)Hours:8 a.m. to 5 p.m. weekdaysContacts:John Strohl, Manager, 515-294-0306 jstrohl@iastate.eduBill Colonna, Fermentation Specialist, 515-294-0643Zhiyou Wen, Professor-in-Charge, 515-294-3011,wenz@iastate.eduLawrence Johnson, CCUR and BCRF Director, 515-294-0160, ljohnson@iastate.eduFax: 515-294-6261 or 515-294-8181 The Fermentation Facility is designed to help researchers develop new fermentation technologies and products and provide benchtop- and pilot-scale fermentation equipment. The facility has equipment for scaling up the production of industrially important chemicals, chemical feedstocks, genetically modified organisms and enzymes.The facility also manages large-scale fermentation activities conducted at the BioCentury Research Farm (BCRF) that are focused on biofuels, industrial chemicals, and biobased products. The Fermentation Facility is a partnership of the Center for Crops Utilization Research,College of Agriculture and Life Sciences, Department of Food Science and Human Nutrition, Department of Chemical and Biological Engineering, and Bioeconomy Institute.Equipment housed in the Food Sciences Building includes four NBS Bioflo 310 benchtop fermentors with 1- and 5-liter working volume capacities and pilot-scale fermenters with 15- to 115-liter working volume capacities. Benchtop fermentors located in Sweeney Hall include two NBS Bioflo 110 units with 1- and 5-liter working volume capacities. Downstream processing equipment includes a hollow fiber ultrafiltration system, semi-continuous centrifuge, French pressure cell press, bio-hood, and an eight-shelf freeze dryer with stoppering capability.Pilot-scale equipment housed in the Biomass Processing Facility at the BCRF, located seven miles west of Ames on U.S. Highway 30, includes two ABEC fermentors with working volumes of 50 and 200 liters, two Applikon fermentors with working volumes of 500 and 1,000 liters, a Sharples P660 continuous horizontal decanting centrifuge, stirred tanks, pumps, and a platform scale. A distillation skid system, falling film evaporator, and rotary drying system have recently been installed and are currently operational.Flow Cytometry FacilityLocation: Main Facility1104 (Office) or 1117 (Lab) Molecular Biology BuildingSatellite Facility1120 Veterinary Medicine ComplexHours:8 a.m. to 5 p.m. weekdays, by appointmentContacts:Shawn Rigby, Manager515-294-2472, rigbysm@iastate.eduChristine Deal, Assistant Scientist515-294-8504Michael Wannemuehler, Professor-in-Charge515-294-3270 Web:biotech.iastate.edu/facilities/cell/The Flow Cytometry Facility of the Office of Biotechnology offers flow cytometric analysis and cell sorting for a wide range of research applications. Facility personnel are trained to assist researchers in flow cytometry experimental design, sample preparation techniques, and data analysis. Training sessions to provide general instruction in these areas are also offered in the facility. All facility services are open to internal Iowa State University clients, as well as to external institutions and individuals.Flow cytometry is used to analyze bacteria, mammalian cells, fungi, yeast, insect cells, microscopic parasites, plant cells, nuclei, organelles and chromosomes. Immunofluorescence measurements are used to provide information on intracellular and cell surface receptor densities. DNA/RNA-specific stains supply information on genome size, chromatin structure, and cell cycle kinetics. Fluorochromes are available for quantitating a number of cellular metabolic functions, such as calcium mobilization and oxidative burst. Phagocytosis of fluorescently labeled particles (beads, yeast or bacteria) can be quantified. Cell viability and concentration can be measured for mammalian cells, as well as bacteria. Fluorescent lipophilic dyes are available that have been utilized as a means of tracking cell life and tissue localization in vivo. Levels of enzyme marker gene expression, such as ?-galactosidase, can be correlated with the fluorescence intensity of cleaved substrate by flow cytometry. Intracellular protein products can be measured by immunofluorescent labeling of fixed cells. Fluorescence in situ hybridization (FISH) techniques provide information on the mRNA expression level of a specific gene and can be used in conjunction with flow cytometry to provide quantitative gene expression information on a cell-by-cell basis. Necrotic versus apoptotic-mediated cell death can be distinguished using flow cytometry. It is also possible to sort individual cell populations via flow cytometry, enabling researchers to separate and further characterize subpopulations of cells.Flow Cytometry Data Acquisition and Cell SortingThe Flow Cytometry Facility maintains three flow cytometers for data acquisition and cell sorting: a BD Biosciences FACSAria III, BD Biosciences FACSCanto and Miltenyi Biotec MACSQuant. The facility also maintains several computer workstations with software packages for performing off-line analysis of flow cytometry data.BD Biosciences FACSAria IIIThe BD FACSAria III is a sorting flow cytometer. The power of multi-color flow cytometry and the highly sensitive analysis platform of this instrument allow investigators to categorize an almost endless variety of unique cell subpopulations. Once cells of interest are identified, the high-speed sorting platform of the FACSAria III can simultaneously isolate and collect these target events for further study.The FACSAria III has three excitation lasers (405, 488 and 633 nm) and is configured to detect nine fluorescent parametes (Pacific Blue, AmCyan, FITC, PE, PE-TexasRed, PerCP-Cy5.5, PE-Cy7, APC and APC-Cy7) as well as forward and side scatter. This gives investigators extensive flexibility in reagent selection and experimental design. The FACSAria III has the capacity for 11-parameter detection on particle sizes ranging from 0.5 to 50 ?m in diameter. Thus, viable cell, aseptic sorts can be performed on a wide range of cell types (i.e. bacteria, mammalian leukocytes, plant protoplasts, etc.). Up to four unique populations can be sorted simultaneously into 1.5 mL microtubes, 12 x 75 mm, or 15 mL tubes. Cells can also be sorted into several different types of tissue culture plates and slides, and both sample and collection chambers can be maintained at pre-determined temperatures.Digital electronics allow the FACSAria III to achieve data acquisition rates as high as 70,000 events per second. For many common sorting experiments, an event rate of 25,000 per second will consistently attain ≥98% sort purity and ≥80% of expected yield. Higher event rate sorts (i.e. 50,000 – 70,000 per sec.) can be achieved in many instances with minimal loss of sort purity and only moderate loss of expected yield.The FACSAria III is located in the Molecular Biology Building (MBB) main facility and is operated by facility personnel only. Data acquired on the FACSAria III is loaded onto a network server which allows customers to retrieve data files for analysis from their office/lab via an Ethernet connection. The Flow Cytometry Facility also maintains a backup of user data. An appointment to schedule an experiment on the FACSAria III can be made by simply calling or emailing Flow Cytometry Facility personnel.BD Biosciences FACSCantoThe FACSCanto is a data acquisition flow cytometer that combines a patented optical design, digital electronics and a novel sample injection system supporting carryover of less than 0.1%. High-speed data processing, industry-leading sensitivity and minimal sample-to-sample carryover make this instrument uniquely suited for rare event analysis. The FACSCanto has the capacity for 10-parameter detection on particle sizes ranging from 0.5 to 50 ?m in diameter, which includes leukocytes, cell lines, platelets, bacteria, multiplexed bead technologies and more.The FACSCanto has two excitation lasers (488 and 633 nm) and is configured to detect eight fluorescent parameters (FITC, PE, PE-TexasRed, PerCP-Cy5.5, PE-Cy7, APC, Alexa Fluor 700, and APC-Cy7), as well as forward and side scatter. Data are acquired with digital electronics. The system can handle high sample flow rates (up to 120 mL/min.) and fast acquisition rates (up 10,000 events per sec.). Digital electronics also facilitate compensation, with no limits to inter- and intra-beam compensation, allowing post-acquisition compensation. Data files are stored raw and compensated as part of the FCS files, allowing flexibility for off-line compensation when viewing data.The FACSCanto instrument is located in the MBB main facility and is operated by facility personnel only. FACSCanto-generated data are loaded onto a network server which allows customers to retrieve data files from their office or lab via an Ethernet connection. The Flow Cytometry Facility also maintains a backup of user data. An appointment to schedule an experiment on the FACSCanto can be made by contacting facility personnel.Miltenyi Biotec MACSQuantThe MACSQuant is a data acquisition flow cytometer that is located in the Veterinary Medicine Complex satellite facility. It is equipped with three excitation lasers (405, 488, and 633 nm) and is configured to detect seven fluorescent parameters (Pacific Blue, FITC, PE, PerCP-Cy5.5, PE-Cy7, APC and APC-Cy7). The MACSQuant is user operated. Required training is available through facility personnel. Once trained, an online calendar allows users to reserve instrument time through any web browser (biotech.iastate.edu/cgi-bin/calweb/calweb.cgi). Archiving MACSQuant-generated data files is the responsibility of individual users.Large-Scale Cell SeparationThe facility maintains an Miltenyi Biotec AutoMACS Pro magnetic cell separation instrument within the Veterinary Medicine Complex satellite office. The AutoMACS Pro is a fully automated bench-top sorter that can be used to perform sterile bulk sorts. Designed for ultra high-speed positive selection as well as depletion, the AutoMACS Pro can isolate virtually any cell type and is compatible with almost any direct or indirect MACS reagent. Users need only label cells to be sorted and choose an AutoMACS Pro separation program. The separation is done automatically. Several separation programs can be selected from a touch screen menu. By using the positive selection program, the AutoMACS Pro is capable of isolating up to 2×108 pure target cells within minutes. Cells as rare as 1 in 106 can be enriched to high purity through the use of double positive selection programs. “Untouched” cells may be obtained just as easily by depleting unwanted cells with the AutoMACS Pro depletion programs. The AutoMACS Pro and associated reagents are completely compatible with flow cytometry. Fluorescent and magnetic labeling of cells can be performed simultaneously. After AutoMACS Pro sorting, cells are immediately ready for flow cytometric analysis. The AutoMACS Pro is user-operated. Potential users must complete a mandatory training session, administrated by facility personnel, which is provided upon request.Additional Laboratory EquipmentRefrigerated table-top centrifuge, carbon dioxide incubators, biohazard hood, temperature-controlled water baths, analytical balances, pH meter, refrigerators, freezers and a cryostorage system are available.Food Science and Human NutritionLocation: 2325 N. Loop Drive, Suite 6100 (ISU Research Park) and 1127 Human Nutritional Sciences BuildingHours: 8 a.m. to 5 p.m., Monday through FridayContacts: Jeanne Stewart, Assistant Scientist II, 515-294-3011, jws1@iastate.eduFax: 515-294-4546The Department of Food Science and Human Nutrition (FSHN) is equipped to facilitate research projects involving human subjects, featuring a variety of research, laboratory space and equipment for controlled studies that involve diet, exercise and behavior change. Equipment may be used by researchers to collect data on metabolic risk factors, body composition and/or physical fitness. An exercise area with private locker room and shower facilities provides convenient space for controlled exercise studies. Exercise equipment includes several treadmills, three cycle ergometers and metabolic carts.Facilities are available for pilot and research studies. Individual testing rooms are available to conduct a variety of clinical measurements. Rooms are set up with metabolic carts and can be used for assessing resting and exercise-related energy expenditure. Another room is designated for body composition via BODPOD, PeaPod, bioelectrical impedance analysis (BIA) and bone mineral densitometry via dual-energy x-ray absorptiometry (DXA). An adjacent room is available for anthropometry assessment. Two phlebotomy rooms are available for collecting blood and other clinical data. A fully-equipped metabolic kitchen with a professional stove/oven, microwave ovens, two dishwashers, refrigerator/freezer, walk-in pantry, walk-in cold room, walk in freezer and an ice machine are available. Adjacent dining rooms provide capacity to serve meals for up to 40 people simultaneously. Additionally, the on-campus facility has a fully equipped kitchen as well as a dining room with the capacity to provide capacity to serve meals for 24-40 participants simultaneously, phlebotomy room and a private room for clinical counseling/interviewing.Estimates regarding costs to conduct human clinical studies are available by contacting the department.Sensory Evaluation Unit (FSHN)Location: 1126 Human Nutritional Sciences BuildingHours:8 a.m. to 5 p.m. weekdays, and as arranged.Contact the facility regarding holiday hours.Contacts:515-294-4323 Ken Prusa, Professor-in-ChargeFax:515-294-2215E-Mail:kprusa@iastate.edu (Ken Prusa) The Sensory Evaluation Unit provides facilities and expertise for the sensory evaluation of food products and/or ingredients.The unit’s facilities include a computerized ten-booth testing area, a panelist training room, a sample preparation area and an adjoining food science and human nutrition laboratory equipped for measuring food color, texture and viscosity.Services include sensory project design and management; panelist recruiting, screening, and training; data collection; graphic presentation of data; and analysis and interpretation of data.Analytical tests available include discrimination tests, descriptive analysis and time intensity. Consumer tests include product preference and acceptance.Fees are based on the actual costs for supplies, labor, the use of the facilities and panelist incentives. For estimates, please contact the facility at 515-294-4323.Genome Informatics FacilityLocation: 206 Science IHours: 8 a.m. to 5 p.m., Monday through FridayContacts: Andrew Severin, Manager515-294-1320, gif@iastate.eduWeb: Genome Informatics Facility of the Iowa State University Office of Biotechnology provides bioinformatics services for investigators within academia, industry and government. Quality analysis starts with understanding the nuances of the biological question and the assumptions made by informatics tools. Facility staff members are available to work closely with researchers to provide the best solution for biological enquiry.Available ServicesGenome assembly and annotationTranscriptome assembly and annotationSNP/InDel callingRNA-seq analysisChIP-seqIntrogression mappingNovel gene discoveryPersonalized GBrowse for data visualizationHigh performance computingTo request bioinformatics support or service, please email the facility at gif@iastate.edu. Include in the email the type of organism and the types of analyses envisioned. An appointment can then be made to further discuss the project.Genomic Technologies FacilityLocation: Center for Plant Genomics2025 Roy J. Carver Co-LaboratoryHours: By appointmentContacts: Patrick S. Schnable, Overall management515-294-0975, schnable@iastate.eduHeng-Cheng “Alvis” Hu, Facility Manager515-294-8563, alvishu@iastate.eduFax: 515-294-5256Web:plantgenomics.iastate.eduThe Genomic Technologies Facility (GTF), a service facility in the Plant Sciences Institute, provides expertise and equipment for biological research at the genomic level. Services include high-throughput genotyping and an optimized maize mutant mapping service (Sequenom-BSA). Next generation sequencing can be performed on the Ion Torrent? Proton or PGM platforms or with the Illumina MiSeq technologies. Two real-time thermocyclers, a Stratagene Mx4000 and a Roche LightCycler 480, are available to internal customers for high-throughput gene expression analysis and single SNP genotyping using Taqman or KASPar markers. The GTF offers Agilent Bioanalyzer, Life Technologies Qubit and E-Gel systems to internal customers for DNA or RNA quantification and fragment size selection. In addition, DNA and RNA extraction services are available.Life Technologies? Sequencing Services (Ion PGM? and Ion Proton?)Ion Torrent? sequencing is suitable for an array of applications that require modest amounts of data generation. Single-end sequencing (100-400bp) is available on the Ion Torrent PGM?. Three chip options are available depending upon the number of reads required: the 314 chip (up to 500,000 reads), the 316 chip (2-3 million reads) or the 318 chip (5-6 million reads). For applications that require more data, 200bp single-end sequencing is available on the Ion Proton using the P1 chip which provides 60-80 million reads per run. Sequencing runs are conducted by GTF staff.Illumina? MiSeq ServicesIllumina?MiSeq sequencing offers an array of applications: resequencing, PCR Amplicon, Library QC and FASTQ-only generation. Paired-end sequencing (2x26bp, 2x75bp, 2x150bp, 2x250bp and 2x300bp) with 12-15 million reads per run is available. Sequencing runs are conducted by GTF staff.Custom NGS Library Preparation ServicesThe GTF offers custom library preparation service for both Illumina and Ion Torrent? sequencing applications.Ion Torrent library preparations available: Ion Xpress Plus Fragment Library and Ion Total RNA-Seq.Illumina also offers serveral library preparation methods, including: TruSeq Small RNA, TruSeq Targeted RNA, TruSeq Stranded Total RNA with Ribo-Zero, TruSeq DNA PCR-Free, TruSeq DNA Nano, TruSeq Custom Amplicon, Nextera XT DNA, Nextera DNA, Nextera Exome Enrichment, and Nextera Rapid Capture Exome and Expanded Exome.Sequenom ServicesThe MassARRAY? system offers a suite of applications for quantitative and qualitative genomic analysis via MALDI-TOF mass spectrometry. A multiplex of up to 40 SNPs in each reaction can be analyzed by iPLEX?, a SNP genotyping technology, providing up to 150,000 genotypes per day. Users provide the sequences of their targets of interest for assay design. Facility staff will confirm the design with users. Assays are conducted by facility staff. Researchers who wish to use an existing set of maize SNP primers developed and validated by the Schnable Laboratory may contact the facility manager.Maize Mutant Mapping ServiceMutants are mapped via Bulk Segregation Analysis (BSA), using Sequenom MassARRAY? iPLEX? technology and over 1,000 validated SNP markers from the Schnable Lab.Before accessing facility equipment and services, users are required to provide a signed and approved user agreement to the facility manager. After training, users can operate some equipment and conduct analyses themselves. In such cases, fees for labor will be applied for the initial training and for experiment preparation. Reservations to use instrumentation may be made using the facility’s on-line scheduler.For related services, please see DNA Facility.Grain Quality LaboratoryLocation:1547 Food Sciences BuildingHours:8 a.m. to 5 p.m. weekdaysContacts:Glen Rippke, Manager, 515-294-5387rippke@iastate.edu,Charles R. Hurburgh, Jr., Professor-in-Charge 515-294-8629, tatry@iastate.eduWeb:The laboratory provides instrumental analyses of the chemical and physical properties of grain and other agricultural products. The services are intended to support high throughput genetic evaluations, surveys of grain quality and other large-scale monitoring operations. The facility also calibrates various types of instruments for manufacturers and users.Chemical composition is measured with a non-destructive, near infrared (NIRS) analyzer. For best accuracy, this test requires 400 grams or more of product. Sample sizes down to 150 grams will work with corresponding reductions in accuracy.Crop-specific tests:Corn: moisture: protein, oil, starch and density; ethanol yield and feed value can be calculated from the NIR resultsSoybeans: moisture, protein, oil, fiber, total saturates and linolenic acid; meal and oil processed value (EPV) can be calculated from the NIR resultsSoybean meal: moisture, protein, oil and fibersDistillers grain: moisture, protein, oil and fiberGeneral TestsSeed weight and sizeBreakage susceptibility (corn)Test weightThins (corn)Specific gravity (measure of hardness) by nitrogen displacementOther contracted servicesAdaptation of NIRS instrumentation for individual situations and specialty measurements is provided upon request.The facility’s calibration service program creates calibrations for electronic measurement equipment of various types and trains users in quality control validation. These can be done for instrument manufacturers or for individual users on a wide range of products and instrument types.The lab has an in-house quality management system to verify accuracy and reproducibility of data. Advice and set-up of quality control programs for analytical data can be provided.Hybridoma FacilityLocation:1106 (Lab) or 1104 (Office) Molecular Biology BuildingHours:8 a.m. to 5 p.m. weekdays, by appointmentContacts:Facility 515-294-9837, hybrid@iastate.eduPaul Kapke, Manager, 515-294-9837Amanda Brockman, Research Associate, 515-294-9837Michael Wannemuehler, Professor-in-Charge, 515-294-3270 Web:biotech.iastate.edu/facilities/hybridoma/The Hybridoma Facility of the Office of Biotechnology provides valuable resources for scientists who need monoclonal or polyclonal antibodies but do not have the appropriate equipment or are not experienced in antibody production techniques. A wide array of procedures can be customized to meet the researcher’s individual requirements. These techniques are provided on an individual charge basis and include animal immunization, cell fusion and hybridoma culture maintenance, cell culture and maintenance of other cell lines used in biotechnology and virology labs, large-scale mammalian cell culture (bioreactor), blood sera collection, antibody purification and isotyping, cryopreservation and cryostorage of cell lines (-140 degrees centigrade), and ELISA tests. The hybridoma projects are usually screened and selected by the client; however, the facility can do the screening and/or training of lab personnel when needed. A hybridoma project usually requires three to five months for completion. The following timetable is used for general planning of a hybridoma project:Mouse immunization—4-6 weeksSelection of primary hybridomas after cell fusion—2 weeksExpansion and freezing of primary hybridomas—2 weeksCloning and screening of clones—2 weeksExpansion and freezing of clones—2 weeksBioreactor/ascites fluid production—4 weeksPolyclonal antibody production in rabbits is available for on-campus clients only. Polyclonal services include the purchase and care of rabbits through the Laboratory Animal Resource group on campus, blood collection and processing of sera, adjuvant addition and injection of antigen and administration of procedures according to an approved protocol and timetable. Additional equipment:9 cubic foot, -80 C freezer2, 18-foot ultra low (cryogenic) freezers for long-term storage of biological materials8 tissue culture incubatorsAll hybridoma services are open to Iowa State University clients and off-campus individuals or companies. The Hybridoma Facility and users will work together to develop an estimate for an individual project based on current pricing. Current price information for each procedure offered may be obtained by calling 515-294-9837 during regular business hours or by sending an e-mail message to hybrid@iastate.edu.Image Analysis FacilityLocation:0119 Molecular Biology BuildingHours:8 a.m. to 5 p.m. weekdays and as arrangedContacts:Margie Carter, Manager 515-294-1011, mcarter@iastate.eduMark Ackermann, Professor-in-Charge515-294-3647, mackerma@iastate.eduThe Image Analysis Facility of the Office of Biotechnology provides 2D and 3D imaging resources for researchers interested in sample measurement or visualization. Services include: 2D image analysis (morphometry, particle analysis, densitometry, etc.), 3D image analysis (volumetric sample measurement) and reconstruction, image editing instruction and photomicroscopy. Analysis is performed with IPLab or Imaris software.Equipment in the facility includes:Imaris for 3D reconstruction and image analysisIPLab for 2D image analysisUpright microscope with fluorescenceInverted microscope with fluorescence and DICAutomated scanning stage with recomposition software for capturing images of large samples at high resolutionStereo microscopeDigital cameras for all microscopes and copy standLaser capture microdissection system to identify and retrieve individual cells from tissue sections. The retrieved cells can be used for assessment/analysis of RNA, DNA, protein and other biochemical properties.Macromolecular X-Ray Crystallography FacilityLocation: 0202 Molecular Biology BuildingHours: 9 a.m. to 5 p.m. weekdays, appointment suggestedContact:James Phillips, Manager, 515-294-2846, jcphill@iastate.eduWeb: Macromolecular X-ray Crystallography Facility of the Office of Biotechnology provides a Rigaku/MSC rotating anode generator for single crystal x-ray diffraction studies, Nikon SMX-800 stereomicroscopes and temperature and vibration controlled chambers for crystallization, training in crystallography techniques, and computers with crystallography software. Facility services are provided on a fee basis and include consultation on protein purification, crystallization, and crystal optimization, as well as assistance with crystal screening, data collection, data processing and structure determination and analysis. The following outline is used for general planning of a crystallography project:Protein purification: concentrated protein > 95% pureCrystallization: 3D, mountable crystalsCrystal screening: search for high quality, high resolution crystalsData collection: using facility or synchrotron equipmentStructure determination: processing, phasing, refinement, modeling, analysis, depositionMaterials Analysis and Research Laboratory Location: 23 Town EngineeringHours: 8 a.m. to 5 p.m. weekdays; Other hours by appointmentContacts: Jerry Amenson, Facility Manager515-294-8752, jamenson@iastate.edu, Fax:515-294-4563Web:marl.iastate.eduThe Materials Analysis and Research Laboratory (MARL) is a core facility of the Office of Biotechnology at Iowa State University. MARL’s function is three-fold, namely research and development, teaching, and service. Its facilities are used for chemical and physical characterization of a wide variety of materials to support research and teaching programs within the university. MARL also conducts research on unusual material evaluation problems for outside agencies.Scanning Electron Microscopy (SEM)The laboratory has two SEMs. The FEI Quanta 250 is a field emission SEM with environmental capabilities (E-SEM). Recent upgrades include a heating stage used to perform dynamic in situ analysis of samples in a temperature range from ambient to 1000°C. The heating stage includes a dedicated gaseous secondary electron detector for sample imaging and recording movies in real time.A beam deceleration option enables adjustment of the landing energy of the electron beam on the sample for conductive samples. This option improves the low kV abberations, resulting in improved imaging at low voltage. In addition, a concentric backscattered (CBS) detector is available to enhance the performance of the beam deceleration option and improve the imaging capabilities at low voltage (<5kV) in the SEM.The other SEM is a Hitachi S-2460N variable-pressure SEM (VP-SEM). Both microscopes have secondary electron and backscattered electron detectors, motorized stages, x-ray analyzers and image analyzers.X-ray Analysis for Microcharacterization (EDS)Both SEMs are equipped with energy-dispersive spectrometers (EDS) for elemental analysis on a microscopic scale. The FEI Quanta 250 is equipped with an Oxford Aztec system, and the Hitachi S-2460N is equipped with an Oxford ISIS system. Both systems have light-element detectors for analysis of elements as light as boron. They have software programs for performing quantitative analysis, digital imaging, line-scan x-ray profiles, multiple element x-ray maps, image analysis and stereo imaging. The system on the FEI is equipped with an 80-mm2 detector to permit good x-ray acquisition, even at low beam currents.X-ray Diffraction (XRD)The lab operates two x-ray diffractometers. One is a Siemens D500 x-ray diffractometer that is equipped with a diffracted beam monochromator and a sample spinner. The diffractometer is fully computer-controlled using hardware and software from MDI, and the user has access to a wide variety of Windows-based data reduction and analysis software. Programs include algorithms for search-matching, quantitative analysis, profile fitting, calculating XRD patterns from known or approximated crystal data, and the determination of crystallite size. Both copper and chromium x-ray tubes are available.The other XRD is a Bruker D8, which is a research grade instrument used to study the underlying structure of solids. The system can be used for phase identification, quantification and structure refinement. The diffractometer is also equipped with a Huber 1/4-cradle so the system can be used for micro-diffraction, residual stress and texture measurements. Two different radiation sources are currently available (copper and chromium) and two different x-ray detectors are available [scintillation detector and a General Area Diffraction Detector System (GADDS)].X-ray Fluorescence (XRF)The lab operates a PANalytical PW2404 x-ray fluorescence spectrometer. The spectrometer is equipped with a 60-specimen sample changer for automated operation. The spectrometer utilizes a rhodium target x-ray tube and a 4kW generator to provide the primary x-ray beam. This provides excellent sensitivity for the determination of all elements from boron through uranium. The spectrometer is fully computer-controlled and the user has access to both qualitative and quantitative analysis software. Specimens can be introduced into the spectrometer as bulk solids (powders, pellets or fused disks) or as liquids or semi-solids. A full range of XRF sample preparation equipment is also available at the lab, including an Eagon 2 automated sample fusion system.Thermal Analysis (TGA and SDT)Thermal analysis is provided with TA Instruments thermogravimetric (TGA) and simultaneous DTA-TGA (SDT) modules. TGA can test samples in inert gases from ambient to 1200°C, with a 25 position autosampler, with a recently-installed Discovery TGA system. SDT can test samples in inert gases from ambient to 1200°C.Image Acquisition, Processing, and AnalysisBoth of the SEM x-ray analyzers are equipped for recording digital images from the SEMs. The FEI SEM has the capability of recording movies. A CCD camera is available for capturing images through the light microscopes or from a camera stand. Noesis Vision’s Visilog software is available for image processing and analysis. A variety of public domain tools for processing and analysis, including Image J, is also available.Light MicroscopyLight microscopes are available for reflected light (Olympus BH), transmitted light (Olympus BH-2) and for stereo (Olympus SZH) imaging. Lenses are available to interface both of the Pixera cameras to the microscopes, for recording still images or movies.Sample PreparationThe lab is equipped with extensive facilities for preparing samples. Equipment includes cutoff saws, grinders, polishers, sputter coaters, evaporators, sample presses, work and Computer SupportMARL operates its own local area network for exchange of data within the laboratory. Results are available for immediate retrieval through WWW and FTP servers.Materials Preparation CenterLocation: Metals Development BuildingHours: 7:30 a.m. to 4 p.m., Monday through FridayContacts: Lawrence L. Jones, Director515-294-5236, jonesll@iastate.eduFax: 515-294-8727Web:mpcThe Materials Preparation Center (MPC) is a U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences, Division of Materials Sciences and Engineering specialized research center located at the Ames Laboratory.MPC is recognized world-wide by the research community for its unique capabilities in preparation, purification, single crystal growth and characterization of rare earth metals, alkaline-earth metals and refractory metal materials. MPC operations are partially supported by the Materials Sciences and Engineering Division as a part of the Synthesis and Processing Science research activity within the Ames Laboratory.Established in 1981, the MPC is a one-of-a-kind facility, which is acutely sensitive to the needs of researchers. Providing research and developmental quantities of high-purity materials and unique characterization services to scientists at university, industry and government facilities on a cost-recovery basis, the MPC allows access to novel materials and new technologies as they are developed.The MPC is renowned for its outstanding technical expertise in alloy preparation, creating materials that exhibit ultra-fine microstructures, high strength and high conductivity – properties of great potential value to American technology. The MPC has established a reputation for close interaction with its clients, providing the kind of personal service required to meet each client’s individual needs. The MPC satisfies requests for customized materials and services that are unavailable from commercial suppliers and unmatched in quality anywhere else in the world. The range of utilization is demonstrated by the scientific publications in which the MPC and Ames Laboratory are cited.Equipment and services information is available on the MPC website.W. M. Keck Metabolomics Research FacilityLocation: 124 Molecular Biology Building (W. M. Keck Laboratory)FT-ICR Mass Spectrometry: 101 Molecular Biology BuildingHours: By appointmentContacts: Ann Perera, Facility Manager515-294-3405Basil Nikolau, Professor-in-Charge515-294-9423 E-Mail:metabolomics@iastate.eduWeb:metabolomics.biotech.iastate.eduThe W. M. Keck Metabolomics Research Laboratory houses seven different analytical platforms, including Agilent gas chromatography instruments with EI and CI ionization methods; an Agilent ion-trap LC/MS instrument with ESI, APCI and APPI ionization capabilities; a Beckman capillary electrophoresis instrument with UV and laser-induced fluorescence; several Agilent 1100 HPLC systems with UV-DAD and ELSD detectors; and a Synergy Two multi-mode microplate reader with the ability to assay optically-active molecules via luminescence, fluorescence, fluorescence polarization, and UV-Vis absorbance (monochromator-based).The laboratory has expanded to include a Bruker Solarix FT-ICR instrument, located in 101 Molecular Biology Building. FT-ICR is a high-end mass spectrometer that provides ultra high-accuracy mass measurements to sub-ppm level.Analysis of samples for external researchers is offered for a fee. After training, internal researchers may choose to analyze their own samples.Microfabrication FacilityLocation:2055 Gilman Hall (Keck Lab)Hours: 8 a.m. to 5 p.m., weekdays; 24-hour access after complete trainingContacts:Krishna Rajan, Oversight515-294-2670, krajan@iastate.edu Fax: 515-294-5444The W. M. Keck Laboratory for the Fabrication of Microminiaturized Analytical Instrumentation (Keck Lab) of the Institute for Combinatorial Discovery provides the ISU and industrial communities with access to state-of-the-art microfabrication technologies. With its approximately 1,000 square feet of class 10/100 clean rooms, the Keck Laboratory supports all phases of microfabrication and its use in fields ranging from analytical chemistry to cell biology. Drawing from affiliates across campus and its resident support staff, expertise in micromechanics, microfluidics, microchip arrays, biology, chemistry, physics and microelectronics can be integrated in translating research ideas into experimental reality.Research capabilities in the laboratory include developing microanalysis systems, chip-scale chromatography, microelectrode assemblies, biochips and cell culture platforms. The laboratory also houses equipment for optical lithography, wet and dry chemical etching and thin film deposition. Analysis and testing equipment, computer workstations and drying and vacuum annealing ovens also are available.Microscopy and NanoImagingFacilityLocation: Room 1, Basement of Bessey HallHours:24-hour access, seven days a week, for individual researchers; professional help available from 8 a.m. to 5 p.m. weekdaysContacts:515-294-3872 Facility, 515-294-1337 (fax)Harry T. (Jack) Horner, Director515-294-8635, hth@iastate.eduTracey M. Pepper, Assistant Scientist and Manager515-294-3872, tpepper@iastate.eduRandall Den Adel, Assistant Scientist515-294-3872, rdenadel@iastate.eduWeb:microscopy.biotech.iastate.edu/The Microscopy and NanoImaging Facility (MNIF) of the Office of Biotechnology provides a variety of instrumentation, technical assistance, consultation and training to individuals and groups of life sciences and biotechnology researchers who want to use photomacrography, light microscopy, scanning and transmission electron microscopy, cryopreservation, cytochemistry, autoradiography, tomography, x-ray microanalysis and image analysis. In addition to the round-the-clock open hours, the MNIF carries out service work for both on- and off-campus researchers. The director and assistant scientist of the MNIF are available for consultation and individual help.Electron MicroscopyElectron microscopy instrumentation includes a 200kV JEOL 2100 scanning/transmission electron microscope (STEM) with elemental analysis, cryo-imaging, tomography, and image analysis systems, and a variable pressure JEOL 5800LV scanning electron microscope (SEM) with an image analysis system.The JEOL 2100 STEM, with <1.4? resolution, consists of the basic STEM, a Noran System 6 light-element energy dispersive x-ray spectrometer (EDS) and integrated software package for computerized control. The digital microscope and analytical system allow elemental analysis of the composition and structure of specimens with a nanometer resolution. Special features include a darkfield/brightfield detector (STEM), hi-angle tilt holders for tomography, cryo-preparation system and holder for cryo-TEM imaging, and two Gatan digital cameras (1K side mount and 2K bottom mount) for image recording, as well as image analysis software.Ancillary equipment available for preparation of specimens for electron microscopy include a Denton DV-502A vacuum evaporator, Denton critical point apparatus, Denton Desk II sputter coater, a Pelco Biowave microwave processing unit, propane jet freezer, two new Reichert UC6 ultramicrotomes, two Reichert ultracut S ultramicrotomes (one with FCS cryo-sectioning system), and a new Leica glass knife maker.The digital JEOL 5800LV SEM with 35? resolution, operates at either high (30-15) or low (10-0.3) kVs and at either high or low pressures to allow for the observation of both fixed and fresh specimens. Image analysis is available for special specimens. Images are captured using the AnalySIS ADDA II digital system with OSIS Pro software.Light MicroscopyLight microscopy instrumentation includes a Zeiss Axioplan II compound microscope equipped with AxioCam color and B/W digital cameras and the following optical modes – bright-field, phase-contrast, polarizing, dark-field, fluorescence and Nomarski (DIC). In addition, this microscope has an Apotome for creating thin optical slices; an Olympus stereomicroscope with reflected and transmitted light sources and a high-resolution digital color camera; dissecting microscopes; a Leitz microspectrophotometer / fluorometer; and Olympus compound microscopes, one with a digital camera. Microscopes with digital cameras have image analysis capabilities.Preparative equipment and space for light microscopy includes fixing and processing hoods; paraffin and resin embedding areas; rotary-, cryo- and ultra-microtomes, knife maker, knife sharpener, Vibratome, microwave, vacuum oven, and centrifuges.Specialized RoomsThe 23-room MNIF, in addition to the microscope and microtomy suites, has two specimen preparation labs; a copy room housing a Bencher photo duplication system and macro photography equipment; a computer suite; a cryoprep lab next to STEM room; an autoradiography/in situ hybridization and developing lab containing an isotope incorporation hood, balance, rotary microtome, tissue culture shaker, refrigerator, oven and processing sink and area; a propane-jet cryopreparation lab with ultralow freezer for substitution; a small prep room with dishwashing sink, deionized water system and autoclave; and a conference room with a library.CryostatAn International Cryostat, Model CTI is available in the MNIF for cryosectioning at temperatures of -18C to -25C. This system uses steel blades and is not automated.InstructionAn Individual Module Training Program (IMTP) was started in Spring 2010 to replace the three graduate-level courses in microscopy that were taught for many years. IMTP trains individuals to specifically process and visualize their research materials using only the methods and instrumentation to accomplish their objectives. This program allows an individual to begin the IMPT within a short time after an initial consultation and signed acceptance of the training conditions. Costs for IMTP include technical training assistance, and instrument, lab and materials fees. The length of an IMPT depends on the individual’s abilities and aptitudes and the complexity of the training. Individuals may contact the MNIF director or supervisor for further details and/or to schedule a consultation.Molecular Printing FacilityLocation:0122 Molecular Biology BuildingHours: 8 a.m. to 5 p.m. weekdays and as arranged; 24-hour access available after complete trainingContact: Curtis Mosher, Manager515-294-6513, cmosher@iastate.eduFax:515-294-7134A new tool for scientific research called the Nano eNablerTM from BioForce Nanosciences, Inc., is available for use in the Roy J. Carver Laboratory for Ultrahigh Resolution Biological Microscopy of the Institute for Combinatorial Discovery.The Nano eNablerTM system is a multifunctional surface patterning platform for dispensing attoliter to femtoliter volumes of biomolecules, nanoparticles and other liquids onto a wide variety of surfaces. The system prints spots and lines from 1 to 60 microns with a 100msec printing cycle, 20 nm stage resolution, 50 mm XY travel and multiplexing ability. This new technology is being applied to a variety of research fields, and the whole spectrum of application of this technology is still being explored. Printable Materials ProteinsQuantum dotsNucleic acidsUV-curable adhesivesLipids EtchantsColloids CatalystsCompatible SurfacesGlass, Silicon, GaN, AlGaN PDMSSilanes HydrogelsMetals NitrocelluloseAlkanethiol monolayers PolystyreneCell PatterningBy patterning substrates with subcellular domains of proteins, it is possible to influence individual cells in a variety of ways. Signaling proteins can be precisely placed and cells grown on the surface contacting deposited materials. Patterning cellular adhesion molecules and blocking the surrounding area make it possible to create arrays of individual or small groups of cells. The highly flexible nature of the instruments allows immediate changes to the patterning of a surface.Biosensor FunctionalizationThe Nano eNablerTM system can be used to functionalize transducing domains in a large variety of biosensors. Small spot sizes mean more testing domains on smaller tests. Small Volume Biomolecular AssaysBiomarker detection and quantification are a good example of small-volume assays with applications including cancer detection, staging, and monitoring, as well as toxicity detection. Plant Transformation FacilityLocation: B423 and B426 Agronomy HallHours: 8 a.m. to 5 p.m. weekdaysContacts:Kan Wang, Director515-294-4429, kanwang@iastate.eduBronwyn Frame, Maize Transformation Team Leader, 515-294-7421 or 8832,bframe@iastate.eduDiane Luth, Soybean Transformation Team Leader515-294-9653, or 4541 dluth@iastate.eduMarcy Main, Rice Transformation, 515-294-6341 or 8832, mmain@iastate.eduKatey Warnberg, 515-294-6341 or 8832, dimmlich@iastate.eduQinglei Gan, Molecular Biology, 515-294-6341 or 5940, qlgan@iastate.eduWeb: agron.iastate.edu/ptf/The ISU Plant Transformation Facility is supported by the Department of Agronomy, the Office of Biotechnology and the Plant Sciences Institute. It offers research partnerships for the genetic transformation of crops. The target crops are corn, soybeans and rice. The facility uses the Bio-Rad Biolistic Apparatus and Agrobacterium tumefaciens as the gene delivery systems for transformation of corn immature zygotic embryos. The Agrobacterium-mediated transformation method is used in the soybean and rice systems.The facility provides expertise in corn, soybean and rice transformation for on- and off-campus researchers. Products provided include transgenic callus, transgenic plantlets and transgenic seeds. The facility also provides a variety of instrumentation, technical assistance, consultation and training to individuals and groups of plant sciences and biotechnology researchers who want to conduct plant transformation. A graduate workshop entitled “Plant Transformation and Transgenic Plant Analysis” (GDCB 542D) is given by the facility every spring semester.Equipment in the facility includes a Bio-Rad Biolistic PDS-1000/He Apparatus, dissecting microscopes, Percival biological incubators, laminar flow hoods, a refrigerated shaker incubator and an SZH10 Olympus fluorescent microscope with digital camera for GFP detection.Protein FacilityLocation: 1178 Molecular Biology BuildingHours: 8 a.m. to 5 p.m., weekdaysContacts: Facility 515-294-3267, protein@iastate.eduJoel Nott, ManagerAmanda Brockman, Assistant Scientist515-294-3267Margie Carter, Assistant Scientist515-294-3267Fax: 515-294-9968 Web:protein.iastate.eduThe Protein Facility provides equipment and expertise for the analysis, characterization and synthesis of proteins and peptides and is open to both internal and external investigators. The Protein Facility is supported by the Office of Biotechnology, the Roy J. Carver Biochemistry, Biophysics and Molecular Biology Department and the Plant Science Institute’s Center for Plant Genomics.The following instrumentation is available in the facility:Amersham Pharmacia DALT 2D electrophoresis systemAmersham Pharmacia Image Scanner with transparency moduleApplied Biosystems Model 494 Procise protein/peptide sequencerApplied Biosystems DE-Pro MALDI mass analyzerApplied Biosystems Q-Star XL quadrapole-TOF tandem mass spectrometer (located in the W. M. Keck Metabolomics Research Laboratory)Three Beckman System Gold high-performance liquid chromatographsModel 125A Analytical Solvent Module with Model 166 programmable UV detectorModel 110A Semi-prep/prep Solvent Module with Model 166 programmable UV detectorModel 125S Microanalytical Solvent Module with Model 166 programmable UV detector and SC100 fraction collectorBeckman Coulter P/ACE MDQ Capillary Electrophoresis SystemBiorad minigel and blotting apparatusC.B.S. Scientific spot-picking workstation with UV lightboxGE Healthcare Ettan IPGPhor 3 IEF unitGenomics Solutions ProGestJasco J-710 SpectropolarimeterMolecular Dynamics Typhoon 9410NanoDrop ND1000 SpectrophotometerSavant SpeedVac PlusVirtis Model 3.5L DBTZL Benchtop FreezeDryer (lyophilizer)Software for the analysis of 1D and 2D gels is also available.The following services are offered.Circular DichroismThe facility provides circular dichroism (CD) spectroscopy as an optical technique to allow the detection and quantitation of the chirality of molecular structures. The CD equipment also is available as a user-operated instrument after a required training session.High Performance Liquid Chromatography (HPLC)The facility offers microanalytical, analytical and preparative HPLC purification of proteins and peptides. The HPLCs also are available as user-operated instruments after a required training session conducted by facility personnel.MALDI-TOF Mass SpectrometryThe facility provides mass spectrometry services for proteins, peptides, glycoproteins, oligosaccharides, oligonucleotides and other polymers using a matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometer. The mass spectrometer also is available as a user-operated instrument after a required training session.Peptide SynthesisThe facility can do both large- and small-scale peptide synthesis, including the synthesis of phosphopeptides, peptides containing unusual amino acids and multiple antigen peptide systems (MAPS) for vaccine production or monoclonal antibody production. The facility also has the capability to synthesize combinatorial peptide libraries.Protein/Peptide SequencingThe facility provides N-terminal protein/peptide sequence analysis of samples in solution or of samples electroblotted onto polyvinylidene difluoride (PVDF) membrane. The facility personnel also performs chemical and enzymatic digestion of proteins in solution or proteins blotted onto PVDF to provide internal sequence information.Q-Star Tandem Mass SpectrometryThe ABI Q-Star XL quadrapole-TOF tandem mass spectrometer is equipped with an oMALDI source, providing a wide range of sensitivity. For both gel-based and chromatographic separations, proteins will be identified, when possible, by searching databases with both MS and MS/MS data using Mascot software. For organisms which do not have complete protein databases, de novo sequences from trypsin fragments can be obtained from MS/MS data. Digestion with other proteases is also available to increase coverage when required. MS-based methods are also available for the identification and mapping of post-translational modification of proteins. A data analysis workstation containing the programs required for the interpretation of mass spectra and database searching is available in the facility. Data analysis can be conducted by individual investigators, with assistance and training from facility staff.In-gel Digestion/Peptide Mass FingerprintingThe facility provides in-gel digestion of protein samples from 1D or 2D gels. Gel spots can be digested with a variety of enzymes including trypsin, Arg-C and Glu-C. The resulting peptides from the digestion can then be analyzed by MS or MS/MS methods. The peptides from the digested proteins can also be separated by HPLC for further analysis by N-terminal sequencing or MALDI-TOF. A UV lightbox and spot picking tools are available for manual gel processing.SDS-PAGE/ElectroblottingThe facility provides SDS-PAGE analysis of proteins for purity and molecular weight estimation and western blotting to nitrocellulose or to PVDF for immuno-detection and protein/peptide sequencing, respectively. Stained gels (Sypro Ruby?, Coomassie Brilliant Blue, silver, etc.) can be scanned and analyzed in the facility. The SDS-PAGE equipment also is available as a user-operated service after a required training session.2-D Gel ElectrophoresisThe facility provides two-dimensional electrophoresis by separating proteins in the first dimension according to charge [isoelectric focusing (IEF)], followed by separating the focused proteins in the second dimension according to molecular weight by SDS-PAGE. The proteins may be visualized by staining with Coomassie Brilliant Blue R250, silver stain or fluorescent dyes. These spots can be excised for further analysis, or the 2D array can be analyzed for differences in protein quantity or in proteins present in the gel. 2D gels also can be electroblotted to PVDF or nitrocellulose membranes for further analysis. The 2D electrophoresis equipment also is available as a user-operated service after a required training session.Isoelectric Focusing (IEF)The facility provides IEF as a method for separating proteins based on isoelectric point prior to SDS-PAGE. The IEF equipment also is available as a user-operated instrument after a required training session, allowing researchers to perform the second dimension in their own labs.2-D Gel AnalysisSameSpots from Nonlinear Dynamics allows for the alignment and analysis of 2-D gels in a short amount of time and returns data that shows the expression of proteins under different biological conditions. The software presents a list of the most significantly changing spots within a set of gels, allowing for the rapid screening of the protein spots that may be of the most interest in a set of gels. These spots can then be further analyzed by other equipment in the Protein Facility (trypsin digestion followed by peptide mass fingerprinting or internal sequencing, direct N-terminal sequencing of the blotted gels or MS/MS using the Q-Star tandem mass spectrometer). This package has the ability to align the gels for complete spot matching. SameSpots is also available for individual use.Image Scanning and AnalysisA Typhoon 9410 Variable Mode Imager is available for scanning of 1D and 2D gels and for phosphor imaging. A screen eraser is available for erasing phosphor imaging screens. The scanner is equipped with three lasers and can scan gels stained with Cy dyes (2D-DIGE gels), Sypro, ProQ and Deep Purple fluorescent gel stains, as well as the common visible stains like silver and Coomassie Blue. The facility also has a flatbed scanner with transparency module for scanning gels and blots. Gel analysis software is available for analyzing 1D and 2D gels, and Amersham’s Decyder Program is available for analysis of DIGE gels. The Typhoon is available for user-based operation as well.qPCR Consultation ServiceLocation: 2736 Veterinary MedicineHours:As arrangedContacts:Jack M. Gallup, 515-294-5844, eag@iastate.eduFax:515-294-5423The qPCR Consultation Service provides in-person and on-line consultation for comprehensive qPCR theory and assay design assistance. All steps are discussed and/or printed out as easy-to-follow processes for immediate in-lab use.Services offered:Basic information/qPCR theory and mathPrimer-probe design assistanceIdentification of appropriate reagents, master mixes and machine platforms for One- and Two-Step qPCR, including LCM-qPCRMIQE-based RNA isolation and other MIQE guideline concerns, DNAse treatment and reverse transcription reaction formulation suggestions and guidanceNucleic acid quality assessment and quantity measurement suggestions and guidanceProcessing of global assay parameters using ISURF software #03407Consultation regarding detection and avoidance of RT and PCR inhibition for all sample types and isolation methodsFile system creation and initial qPCR test plate set-up printouts and consultationProcessing of test plate results into final set-up parameters and procedural printouts for final sample qPCRExcel spreadsheets custom-created for EAMP-corrected data analysis and graphingAll plant and animal species are considered. For more information regarding fees and services, contact the manager.Roy J. Carver Laboratory for Ultrahigh Resolution Biological MicroscopyLocation: 0122 Molecular Biology BuildingHours:8 a.m. to 5 p.m. weekdays, and as arranged 24-hour access after complete trainingContacts:Curtis Mosher, Manager, 515-294-6513, cmosher@iastate.eduFax:515-294-7134The Roy J. Carver Laboratory for Ultrahigh Resolution Biological Microscopy located in the Molecular Biology Building provides an atmosphere for the development of interdisciplinary research between the life and physical sciences. Researchers can use the instrumentation housed in the laboratory to perform optical and scanning force imaging of their samples. Lab space is available for students working on projects in the laboratory. In addition, the Carver Lab has a fully equipped cell culture facility and a data processing center where experiments can be analyzed and the results copied onto CDs for use in researchers’ home laboratories.See also Atomic Force MicroscopyOptical WorkstationThe workstation consists of an inverted Nikon Eclipse 200 microscope with bright-field and differential interference contrast optics, fluorescence and wavelength separation to two digital cameras. Software controlling the optical workstation is designed to automate image acquisition. The fluorescent system on the workstation has filter sets for DAPI, FITC, CY3/TRITC, CY5, GFP, CFY and YFP. Images are stored on a hard drive and are available for export via the Internet and CD. The Nikon Eclipse is equipped with high numerical aperture objectives from 2X to 100X.Hyperspectral WorkstationThe latest addition to the laboratory is an Optical Insights hyperspectral microscope. Designed specifically to eliminate problems inherent in fluorescence microscopy, this system allows spectral separation of defined objects in the data set of imaged specimens. This workstation consists of the Optical Insights instrument attached to an inverted Nikon Eclipse 2000 equipped with a Photometrics Cascade 512B digital camera. The system has a wide range of customized fluorescent filter cubes and microscope objectives. This system has software specific for hyperspectral data acquisition and analysis, in addition to the latest MetaMorph (7.0) imaging software which allows 4D viewing and 3D measurements.Additional hyperspectral and MetaMorph analysis software is installed on computers in the laboratory’s data processing room. ................
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