UNC Biochemistry Core Facilities and Resources

 

 

 

These pages list our core facilities by areas of focus.  These are brief profiles of the cores; to find more detailed information on each core, including location, contact information, and equipment lists, please check out their facility overview in their listing in our searchable database.

 

The biochemistry cores offer resources to investigators working on the molecular analysis of genes, proteins, peptides, cells and tissues.  Analytical approaches include mass spectrometry, NMR spectroscopy, x-ray crystallography and bioinformatics.

Immunology


CFAR Virology, Immunology & Microbiology Core 
provides support for research among CFAR (Center for AIDS Research) investigators and training in sample preparation and/or the performance of immunologic procedures.

Research applications: Sample processing and storage, HLA-typing, CD4 counts, measurement of soluble biomarkers (ELISA, Luminex platform, ELISPOT), flow cytometry for phenotyping and functional analysis of immune cell populations; antibody analysis.

Instrumentation available: Luminex, MagPix Instruments, 6-8 color flow cytometry instruments –FACS Canto (BD), FACS Calibur (BD), ELISA reader, multiple PCR instruments.

Eligible Users: UNC infectious disease clinicians, CFAR–affiliated researchers, biotech companies, other clinicians and researchers interested in infectious diseases.

 

The Antibody Core Facility can help investigators produce monoclonal antibody-producing hybridoma cell lines. The core has partnered with an outside vendor (Precision Antibody) to work with investigator labs to develop hybridoma lines. Once developed, the core can take possession of these lines, store them for the investigators, and carry out large scale production and purification of the antibodies (multi-milligram). The facility also can produce large amounts of crude antibody in vitro for investigators (conditioned media) and purifies both polyclonal and monoclonal antibodies from supplied tissue culture media or ascites. If the Antibody Core is able to extract the antibody sequence from a hybridoma cell line, the Protein Expression and Purification Core – located in the same building – can produce it recombinantly using mammalian expression systems. This facility can also help you to produce protein antigen(s) you may require for your hybridoma project.

Operation: We help the investigator lab think through and structure their project, coordinate its execution and produce the resulting custom antibodies in our facility once the hybridoma lines are established. We help them produce antigens as needed via the Protein Expression and Purification Core, connect them with our commercial partner precision antibody, and try to facilitate the process in any way we can

Research applications: Generation of hybridoma cell lines from mice, rats and hamsters to produce monoclonal antibodies that specifically recognize an antigen(s) of interest. Design and production of protein antigens in the Protein Expression and Purification Core, which is located in the same facility. Production of antibody from of hybridoma cell culture. Purification of polyclonal and monoclonal antibodies from tissue culture media or ascites. Conjugation of purified antibodies to a variety of things like fluorescent tags, small molecules like biotin, other proteins, and chromatography media. Tissue culture services are also offered- examples: adapting cell lines to various medias and suspension culture.

Instrumentation available: Animal work is done with our commercial partner (Precision Antibody). Notable equipment – a Biotek plate washing robot which enables high throughput ELISA screening; several western blot devices that enable screening of up to 50 sera per blot. Copious amounts of 37C CO2 regulated incubator space (including shaking platforms). We also are co-localized with the protein expression and purification core, providing access to all their resources.

Training resources: Individualized training (fee-for-service) of users for some aspects of our service is offered, please contact for more detail.

Eligible users: Researchers at UNC, other academic institutions as well as industrial laboratories.

Mass Spectrometry


The Biomarker Mass Spectrometry Facility provides expertise for qualitative and quantitative analysis of small molecular weight biomarkers including nucleic acids, peptides, and metabolites. In addition to LC/MS/MS analyses, we offer HPLC with UV detection and trace elemental analysis by ICP-MS. Areas of focus for the facility have included DNA and protein adducts as markers of chemical exposure, oxidative stress, and endogenous DNA damage, characterization of chemical reaction products, identification and quantitation of metabolites, multi-element analysis and arsenic speciation. The services and analyses performed within the facility enable UNC investigators to study the molecular mechanisms of environmentally based diseases as well as the relationships between genetic and environmental factors.

Research applications: Targeted quantitation of biomarkers of exposure and effect including DNA adducts, protein adducts, DNA-DNA and DNA-protein crosslinks, metabolites, and metals; Unbiased differential mass profiling; HPLC purification of reaction mixtures; Molecular weight and structural characterization of synthetic standards.

Instrumentation: Sciex 6500 triple quad mass spec with Eksigent nanoLC; Thermo TSQ Quantum Ultra triple quad mass spec with Waters NanoAcquity UPLC; Thermo TSQ Quantum Ultra triple quad mass spec with Waters Acquity UPLC; Agilent 6520 Accurate-Mass Quadrupole Time-of-Flight mass spec with Agilent 1200 Rapid Resolution LC; Agilent 7500cx ICP-MS Inductively Coupled Plasma mass spec with Agilent 1260 Infinity Bioinert HPLC; Agilent 1200 HPLC for offline purification and fraction collection.

Resources: Full service sample analyses as well as training for independent operation; Consultation for proposal submission and/or project feasibility; Method development; Data interpretation and generation of experimental sections and figures for publications. Fees are based on hourly usage of instrumentation and staff time, so costs correlate with the degree of effort required and the duration of analyses.

Eligible Users include UNC researchers as well as other academic investigators and commercial clients when feasible.

The Department of Chemistry Mass Spectrometry Core Laboratory specializes in small molecule mass spectrometry analysis. Our Core aims to provide the highest quality mass spectrometric services possible to both campus and surrounding Research Triangle area members. We offer both independent operator and full-service mass spectrometry workflows. Our primary research focuses are organic and inorganic synthetic molecules, relative and absolute quantitation mass spectrometry, and trace metal analysis.

Research applications: quantitative mass spectrometry, liquid chromatography separations, structural elucidation, MS/MS & MSn fragmentation mass spectrometry, complex mixture analysis, molecular formula confirmation, high resolution and accurate mass analysis, & trace metal determination/quantitation.

Instrumentation available: ThermoFisher Exactive GC, ThermoFisher LTQ FT with a Waters Acquity H-Class UPLC system, PESciex 3000 with a Shimadzu LC and CTC autosampler, ThermoFisher Element XR equipped with ESI Autosampler and Teledyne Cetac Excite 193 Laser Ablation system, Agilent 1100 liquid chromatograph system, CEM Mars Microwave System

Resources: Individualized user training for undergraduate, graduate, and post-doctoral scholars of the Department of Chemistry and UNC Campus community. Walk-up access is available to all trained users. Full-service sample analysis. Consultation for project development. Method development and advising. Data interpretation. Grant application assistance.

Eligible users: All researchers at UNC-CH, affiliated universities, non-affiliated universities, private industry/commercial clients.


UNC Michael Hooker Proteomics Center provides researchers with state-of-the-art analysis of proteins from tissues, cells and other biological samples. The Center is also a resource for hands-on-training, education, and consultation in modern proteomics techniques. The Center offers an array of sample preparation and instrumentation services, as well as method development and in-depth consultation to help researchers optimize their experimental design.

Research applications: Identify purified proteins and co-immunoprecipitated proteins. Identify proteins in from complex mixtures and quantify differences in protein abundance using labeled (SILAC, TMT) or label-free approaches. Identify of post-translational modifications such as acetylation, ubiquitylation, phosphorylation and sulfhydryl modifications. Determine the exact mass of purified peptides and proteins.

Instrumentation available: Thermo Easy nLC 1000; Thermo QExactive HF mass spectrometer; Waters nanoAcquity; Thermo LTQ-Orbitrap Velos mass spectrometer; Sciex 5800 MALDI-TOF/TOF mass spectrometer

Training resources: Individualized training is available for the use of the MALDI-TOF/TOF instrument and data analysis using Protein PilotTM software.

Eligible users: Researchers at UNC and other academic institutions or industrial laboratories.

Metabolomics


The Metabolomics Laboratory is a collaborative resource for global and targeted metabolite profiling from a wide range of biological samples.

Research applications: NMR analysis and absolute quantitation on nearly 300 metabolites including, amino acids, organic acids, amines, nucleic acids, carbohydrates, drug related metabolites, etc. from a variety of biological samples (such as urine, serum, CSF and saliva) cell and tissue extracts. Based on research needs, perfused bioreactor studies are available for intact tissue samples or cell suspensions. Concentrations ranging from micromolar up through millimolar levels can be detected. A microcoil NMR probe requiring only 15 µl injections that can be recaptured after analysis, permits concentration of samples thereby pushing detection limits to low micromolar levels, with samples amounts of just 5 µl - typical of mouse plasma metabolomic studies. Typical output for users is a principal component analysis of PC1 versus PC2 with a list of biomarkers derived from the loadings plot.  For <15 media, plasma, or urine samples plan for a week turn around.

Instrumentation available: Varian INOVA 600MHz NMR 
2 channels with pulsed field gradients, and waveform generators, used with Varian 5mm PFG ID probe
or Protasis 10µl HC capillary NMR probe. A Varian SMS 100 robotic sample changer for can queue up 100 samples in 5mm NMR tubes for automated analysis. A Protasis High Throughput fluidics system for the automated injection of small samples (down to 15 µl volume) has three temperature controlled drawers and can accommodate up to 2 x 96 well-plates or 200 of vials per drawer (600 samples total) for automated fluidic injection of samples into the Protasis 10µl HX capillary NMR probe.

Eligible users: Researchers at UNC and other academic institutions or industrial laboratories.

Nuclear Magnetic Resonance (NMR)


The UNC Biomolecular NMR Laboratory supports research into the structure and dynamics of biological molecules.

Research applications: Macromolecular structure and dynamics. Cryoprobes are available for small molecules and biological applications, including small molecule and natural product NMR.

Instrumentation available: Bruker Avance 500 MHz with cryoprobe, Bruker Avance 600 MHz with cryoprobe, Bruker Avance 700 MHz with coldprobe and SampleJet 500 slot sample changer, Bruker Avance 850 with cryoprobe and 24 slot sample changer.

Training resources: one-on-one training of new users, consultation with prospective users to determine whether NMR will be useful in their research, and consultation in designing experiments. NMR course BIOC663 open to graduate students.

Eligible users: Any researchers at UNC or other academic institutions, and in industry.


The Eshelman School of Pharmacy NMR Facility
provides a resource for small molecule and natural product NMR related to drug discovery and can also be used to study polymers and biomolecular structure and dynamics.

 Research applications: Small molecule and natural product NMR; polymer NMR; biomolecular structure and dynamics. In addition to 1H, 13C, 31P, and 19F, we have studied more unusual nuclei, such as 11B, 129Xe, and 119Sn.

Instrumentation available: Fully automated Varian 400MR used for 1H, 13C, 31P, and 19F one-dimensional experiments, as well as those two-dimensional experiments often associated with “small” molecules, such as COSY (and its variants), NOESY, ROESY, HSQC, HMBC. The Inova 400 is used for 1H and 13C one-dimensional experiments, and for the same sort of two-dimensional experiments as are run on the 400MR. The Inova 500 was originally designed for biomolecular NMR, and so is our most versatile instrument, with 4 RF channels instead of 2, all equipped with wave-form generators; triple-axis gradients; pre-cooling of the VT air so as to allow temperature studies below room temperature; and a variety of probes, which among other things allow us to study not only 1H, 13C, and 19F, but also 31P, 11B, 129Xe, and 119Sn.

Training resources: Frequent training sessions (from 1-4 trainees) scheduled based on demand. Free consultations for prospective users.

Eligible users: Any researchers at UNC or other academic institutions, or industry.

 

The Metabolomics Laboratory (by NMR) is a collaborative resource for global and targeted metabolite profiling from a wide range of biological samples.

Research applications: NMR analysis and absolute quantitation on nearly 300 metabolites including, amino acids, organic acids, amines, nucleic acids, carbohydrates, drug related metabolites, etc. from a variety of biological samples (such as urine, serum, CSF and saliva) cell and tissue extracts. Based on research needs, perfused bioreactor studies are available for intact tissue samples or cell suspensions. Concentrations ranging from micromolar up through millimolar levels can be detected. A microcoil NMR probe requiring only 15 µl injections that can be recaptured after analysis, permits concentration of samples thereby pushing detection limits to low micromolar levels, with samples amounts of just 5 µl - typical of mouse plasma metabolomic studies. Typical output for users is a principal component analysis of PC1 versus PC2 with a list of biomarkers derived from the loadings plot.  For <15 media, plasma, or urine samples plan for a week turn around.

Instrumentation available: Varian INOVA 600MHz NMR 
2 channels with pulsed field gradients, and waveform generators, used with Varian 5mm PFG ID probe
or Protasis 10µl HC capillary NMR probe. A Varian SMS 100 robotic sample changer for can queue up 100 samples in 5mm NMR tubes for automated analysis. A Protasis High Throughput fluidics system for the automated injection of small samples (down to 15 µl volume) has three temperature controlled drawers and can accommodate up to 2 x 96 well-plates or 200 of vials per drawer (600 samples total) for automated fluidic injection of samples into the Protasis 10µl HX capillary NMR probe.

Eligible users: Researchers at UNC and other academic institutions or industrial laboratories.

Peptide Synthesis


High Throughput Peptide Synthesis and Array Facility
provides researchers with high quality consultation and peptide synthesis services including design, production, purification, and characterization of peptides and custom designed biotinylated peptide arrays.

Research applications: Synthetic peptides containing standard, post-translationally modified (e.g., acetylation, methylation, phosphorylation) and/or unnatural/nonstandard amino acids and fluorescent tags (e.g., 6-Cl-Trp, FAM-, TAMRA-) can be synthesized for a wide range of biological and biophysical applications, including FP, ITC, NMR, X-ray crystallography, mass spectrometry (including stable isotopes containing peptides, with isotopes such as 2H, 13C, and 15N), enzyme inhibition/competition assays, protein-protein interaction studies, cell permeable peptide reagents, antigen/antibody production including synthesis of Multiple Antigenic Peptides (MAPs), and high density peptide arrays.

Instrumentation available: Symphony multiple peptide synthesizer, PS-3 peptide synthesizer, Waters preparative, semi-preparative and analytical HPLC instruments, OmniGrid 100 microarray printer. Freeze dryers and a microbalance with a static charges neutralizing device.

Training resources: Consultation with users to determine whether synthetic peptides and peptide chemistry may be useful in their research, and consultation in designing peptides optimal for their experiments.

Eligible users: Any researcher at UNC, other academic institutions, and industrial laboratories.

 

Structural Biology


The Macromolecular Interactions Facility (Mac-In-Fac) provides instrumentation and resources for characterization of biological macromolecules and their interactions with cognate ligands.

Research applications: Determination of solution molecular weight, oligomeric state and association/dissociation constants of macromolecules; on and off rates (kon and koff), stoichiometry (N), equilibrium binding constant (Kd) and other thermodynamic parameters (∆G, ∆H and ∆S) of bimolecular interactions; secondary structure, melting profile, melting temperature (Tm), conformational and colloidal stability, sedimentation, diffusion and frictional coefficients, hydrodynamic and RMS radius, and second virial coefficient of macromolecules.

Instrumentation available: Isothermal Titration Calorimeters (Malvern VP-ITC and the fully-automated Auto-ITC 200); Surface Plasmon Resonance (SPR)-based biosensor (Bio-Rad Proteon XPR36); Bilayer Interferometry (BLI)-based biosensor (ForteBio Octet Red384); Circular Dichroism spectropolarimeters (Applied Photophysics Chirascan-Plus); Size Excusion Column with Multi-angle Light Scattering System (Wyatt DAWN Heleos II); Dynamic Light Scattering instrument (Wyatt DynaPro platereader); Analytical Ultracentrifuges with absorbance and interference optics ( Beckman Optima XL-I); Fluorescence microplate reader with built-in reagent injector (BMG-LABTECH Pherastar FS); T-format Spectrofluorometer (Horiba Jobin Yvon Fluorolog 322); Differential Scanning Fluorimeter (Nanotemper Prometheus NT.48); Micro Scale Thermophoresis (MST) instrument (Monolth NT.115); and Differential scanning calorimeter ( Malvern VP-DSC).

Training resources: One-on-one training of new users, consultation with prospective users to determine whether techniques available at the Mac-In-Fac will be useful in their research, and consultation in experimental design, data analysis and interpretation. The Macromolecular Interaction course BIOC662 taught by Ash Tripathy (Director of the Facility) each spring, is open to graduate students. The course includes both theory and practicals.

Eligible users: Any researchers at UNC or other academic institutions, and in industry.

 
Macromolecular X-Ray Crystallography Core Facility 
(MX) provides support and infrastructure necessary to enable individual investigators of any experience level to initiate and complete a macromolecular structural biology project.

Operation: Three general ways users interact with us that can be adjusted at any time: (1) Facility can be an extension of your lab, with trained users operating independently. (2) We collaborate with your group, mentoring your personnel, helping with experimental design and analysis, and aiding with grant writing and manuscript preparation (3) We independently tackle your project.

Research applications: High-Throughput protein crystallization screening, monitoring, and crystal diffraction testing as well as data collection, crystal structure determination and analysis. Sample preparation may be done in the Protein Expression and Purification Core, which is located in the same facility.

Instrumentation available: Crystallization - Wyatt plate based (384) DLS instrument for validation of sample quality; Rigaku Phoenix and Douglass Instruments Oryx4 liquid handling robots for 96 well crystallization plate setup; an crystallization robot for an Alchemist II liquid handler for custom 96 and 24 well crystallization screen generation; a Minstrel HT imager for automated monitoring of crystallization trials; Jansi UVEX UV florescence microscope for inspection of crystallization experiments as well as multiple Leica stereo light microscopes with cameras for manual plate inspection and crystal harvesting/manipulation. Diffraction - Rigaku Micromax 007HF generator: Port one equipped with an R-Axis IV++ detector optimized for crystals with a long cell axis; Port two is equipped for automated diffraction screening with an ACTOR robotic system and Saturn 944+ CCD detector. Both beamlines are equipped with an X-stream 2000, enabling cryo-crystallography. Access to synchrotron Beamtime at the Advanced Photon Source at Argonne National Lab in Chicago; UNC is a member of the South Eastern Collaborative Access Team (SER-CAT)

Training resources: Individualized training (fee-for-service) of users in any aspect of a crystallography project.

Eligible users: Researchers at UNC, other academic institutions as well as industrial laboratories; UNC LCCC members receive discounted fees for certain services.

 
The Protein Expression and Purification Core (PEP) specializes in the production of pure, functional proteins for structural, biophysical, and biochemical studies. It is specifically designed as a “front-end” interface to other components of the UNC Center for Structural Biology (CSB). In addition, the services of this core are also applicable to other research areas, including recombinant antibody and antigen production as well as stable cell line generation for protein production.

Research applications: Production of purified protein suitable for biochemical, biophysical, and structural studies, the latter of which may be completed in the Macromolecular x-ray Crystallography Core located in the same facility. Produce purified protein suitable for antibody production, which may be done in the Antibody Core Facility located in the same facility. Generation of stably transfected, expression optimized mammalian cell lines. Their uses include: (1) production of your protein of interest in high amounts over multi-week timescales. (2) The cells themselves are a reagent for certain cell based assays.

Instrumentation available: The Core possesses the resources needed for growth and maintenance of large volumes of E. coli (temp. controlled shakers and a 10L fermenter), insect cells (baculovirus mediated, Sf9 and Hi-five cell lines), and mammalian cells (including HEK 293s and CHOs, both suspension and adherent.). A variety of FPLC systems for automated, multi-step, multi-sample purifications as well as more traditional single step, single sample purifications and method development. We also have equipment that enables gel electrophoresis/western blotting and imaging, small-volume (.5ml to 400ml) as well as large volume (<10L) sample concentration, spectrophotometric analysis (NanoDrop), PCR, cloning, electroporation of E. coli and mammalian cells, as well as limited proteolysis analysis of your protein sample.

 Training resources: Individualized training (fee-for-service) in methods of protein expression and purification as well as instrumentation operation.

Eligible users: Researchers at UNC, other academic institutions as well as industrial laboratories; UNC LCCC members receive discounted fees for certain services.

The R. L. Juliano Structural Bioinformatics Core Facility provides researchers with state-of-the-art consultation and collaboration in research studies requiring computational structural biology methods.

Research applications: Bioinformatics and Molecular Evolution analyses include multiple sequence alignments, phylogenetic analysis, and ancestral sequence predictions. Protein structure analysis includes protein fold determination, structure 
superimposition and comparison, homology modeling, active site identification, prediction of protein-protein interaction interfaces, mutation effects analysis, electrostatic surfaces, solvent accessibility calculations, subunit contact analysis, molecular dynamics calculations, free energies of binding, and docking of small molecules.

Instrumentation available: Researchers utilize hardware and software resources provided by ITS Research Computing. Available software packages include but are not limited to PyMOL, BLAST, T-Coffee, Discovery Studio, VMD, GROMACS and Amber.

Training resources: Training workshops on the following topics are available upon request: (1) Protein Data Bank (PDB), (2) PyMOL, and (3) Protein fold recognition and homology modeling. Individualized consultation and collaboration on research studies requiring computational structural biology methods is also available.

Eligible users: Researchers at UNC and other academic institutions or industrial laboratories.

Protein Expression

The Protein Expression and Purification Core (PEP) specializes in the production of pure, functional proteins for structural, biophysical, and biochemical studies.  Services of this core are applicable to other research areas, including recombinant antibody and antigen production as well as stable cell line generation for protein production.

Research applications: Production of purified protein suitable for biochemical, biophysical, and structural studies, the latter of which may be completed in the Macromolecular x-ray Crystallography Core located in the same facility. Produce purified protein suitable for antibody production, which may be done in the Antibody Core Facility located in the same facility. Generation of stably transfected, expression optimized mammalian cell lines. Their uses include: (1) production of your protein of interest in high amounts over multi-week timescales. (2) The cells themselves are a reagent for certain cell based assays.

Instrumentation available: The Core possesses the resources needed for growth and maintenance of large volumes of E. coli (temp. controlled shakers and a 10L fermenter), insect cells (baculovirus mediated, Sf9 and Hi-five cell lines), and mammalian cells (including HEK 293s and CHOs, both suspension and adherent.). A variety of FPLC systems for automated, multi-step, multi-sample purifications as well as more traditional single step, single sample purifications and method development. We also have equipment that enables gel electrophoresis/western blotting and imaging, small-volume (.5ml to 400ml) as well as large volume (<10L) sample concentration, spectrophotometric analysis (NanoDrop), PCR, cloning, electroporation of E. coli and mammalian cells, as well as limited proteolysis analysis of your protein sample.

Training resources: Individualized training (fee-for-service) in methods of protein expression and purification as well as instrumentation operation.

Eligible users: Researchers at UNC, other academic institutions as well as industrial laboratories; UNC LCCC members receive discounted fees for certain services.

 

Analytical and Physical Chemistry

The Nanomedicines Characterization Core Facility (NCore) provides comprehensive characterization of novel nanomedicines and nanomaterials. Services include physicochemical assays (ICP-MS, Nanosight, Capillary Electrophoresis, WES Simple Western Blotting, HPLC, FPLC, Thermogravimetric Analysis, GPC, Malvern Zetasizer, and more). The Nanomedicines Characterization Core Facility (NCore) grants researchers an opportunity for in-depth characterization of nanomaterials and carrier-mediated agents (CMA) including: chemical composition, number average molar mass Mn, weight average molar mass Mw, polydispersity, molecular weight distribution, size, gyration radius, size distribution, shape, surface charge, count number of nanoparticles, solubility (second virial coefficient), thermal/oxidative stabilities, estimation of product lifetime, isotope analysis, analytical evaluation of nanoparticle encapsulated and released drug or agent stability and released rates, degree and rate of recognition of nanoparticles by mononuclear phagocyte system (MPS), UV/Vis spectroscopy, UV-temperature correlation, etc.

 Research applications: Physicochemical characterization (PC) of nanomaterials offers a comprehensive body of information that can be used to predict behavior of the nanomaterials in biological environments both in vitro and in vivo. This information is essential in order to exercise tight control over inter-batch variations and maintain reproducibility of nanomaterials critical for their applications.

 N-Core offers PC of various classes of nanomaterials such as:

  • Polymer conjugates
  • Polymeric micelles
  • Liposomes
  • Nanogels
  • Polyion complexes of small drugs and biomacromolecules (proteins, DNA, and RNA)
  • Inorganic/metal nanoparticles
  • Bio-derived nanoparticles such as exosomes with protein and nucleic acid car

 Eligible users: Any researcher at UNC or other academic institutions, and industry.

 

The CGIBD Advanced Analytics (AA) Core  provides three main services: 1) single-cell genomic and proteomic analysis, 2) high-throughput qPCR, and 3) cost-effective immunoassays. 

Operation: The AA Core offers turn-key research solutions – you bring us samples, we send you processed data. Each project begins with an initial consultation to discuss experimental design and feasibility. For qPCR and immunoassays, we’ll also discuss commercially available assays suitable for your application and obtain any kits or reagents required for the services you need. For single-cell analyses, we’ll schedule a subsequent experiment date (or dates) on which you’ll provide a freshly isolated and filtered cell suspension, after which the Core will handle the rest: FACS isolation of single cells, live cell capture, cDNA library prep, QC, and any downstream processing required (including Illumina library prep, gene expression analysis, sequencing, and data processing via customized pipelines). Cells used for single-cell sequencing can also be applied to single-cell Western blotting in order to interrogate the expression of key proteins identified in the sequencing data. Users receive their results electronically after work has been completed and can track the progress of their projects via cloud-based reports. The AA Core also offers assistance with end-user data analysis.

Research applications:
Single-cell analysis
: single-cell mRNASeq, DNASeq, ATACSeq, specific target amplification (STA), and Western blotting. Both Fluidigm and DropSeq-based technologies are available for single-cell RNASeq, and can be used in tandem.

High-throughput qPCR: gene expression (GE) analysis, SNP genotyping, copy number variation, and quantitative digital PCR, with up to 9216 GE/SNP qRT-PCR or 36960 qdPCR values per run.

Cost-effective immunoassays: Luminex bead-based multiplex assays, ELISA, and DropArray “mini”-multiplexes, which use 80% less sample and reagent volume.

Instrumentation available: Bio-Rad/Illumina ddSEQ Single-Cell Isolator, ProteinSimple Milo Single-Cell Western Blot, Fluidigm C1 Single-Cell Autoprep System, Fluidigm BioMark HD High-Throughput qPCR, Sony SH800Z 6-color FACS, Qubit 3.0 fluorometer, Bio-Rad Bio-Plex 200 bead-based immunoassay system, Curiox DropArray multiplex miniaturization system, BioTek Synergy HT multi-mode microplate reader, and a BioTek ELx405rs microplate washer; the AA Core also has access to an Agilent Bioanalyzer, Agilent TapeStation, and an Illumina NextSeq 500 through arrangements with other labs/core facilities.

Training resources: Free consultation for all prospective users. Instrument training is available upon request to CGIBD members for certain instruments and applications.

Eligible users:  All UNC investigators, as well as PIs from other academic institutions and corporate/industry labs.

 

 

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