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Technologies/Expertise

The technologies and expertise of the four laboratories/groups listed immediately below can be viewed via their individual Web pages:

You may search the rest of the Advance Technology Program’s technologies and expertise by entering a term in the field below and clicking on the "Search" button. If you would like to sort the technologies list by lab, click on the "Show Labs" link located below the search area.

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Amino Acid Microsequencing
  Preparation of protein samples for amino acid analysis
  Protein microsequencing (Edman degradation)
Biocore® Analysis of Macromolecular Interactions
  Data analysis
  Macromolecular interactions: DNA-protein, protein-protein, protein-ligand interactions
  Molecular mechanism-based small molecule inhibitor screen
Capillary Electrophoresis (CE)
  Determination of protein/drug or protein/DNA interactions
  Fast separation of DNA fragments and PCR products
  Purity determination of proteins, peptides, and monoclonal antibodies
  Ultrasensitive laser-induced fluorescence detection
Cell-Free Expression
  Expression of proteins using cell extracts
  Improved solubility of intractable proteins
Clinical Diagnostics
  Mycoplasma testing
  Pathogen identification and classification
  SNP genotyping
Confocal Microscopy
  Auto-archiving of all images; users access their images via the Internet
  Bright-field, phase, and differential interference contrast (Nomarsky) imaging in conjunction with fluorescence imaging
  Experiments involving the uncaging of molecules in cells
  FRAP (fluorescence recovery after photobleaching) – Measures the diffusion of fluorescence-tagged molecules. First, the fluorescent molecules are photobleached in a defined region of the cell. Next, we observe the recovery of fluorescence into this same region from the unbleached surroundings.
  FRET (fluorescence resonance energy transfer) - Detects the binding of two fluorescence-tagged molecules in cells.
  Multi-color, high-resolution fluorescence image acquisition of fixed and live specimens (up to 3 colors). Specimens can be up to 50-microns thick.
  Ratio imaging of calcium ions and other elements
  Second-harmonic generation imaging microscopy (SHIM)
  Time-lapse imaging of live cells
Data Analysis/Management
  Laboratory Information management systems
  Specialized data/database analysis software
DNA Sequencing
  Individual samples and high-throughput
  Next generation 454 high throughput sequencing
  Re-sequencing/mutation detection
DNA/RNA Real Time Quantitative Expertise
  CDNA synthesis
  Data analysis
  Establishment of dual labeled probes
  Establishment of endogenous controls
  Real-Time PCR assay design and development
  Set up and run assay
  Total or cytoplasmic RNA and poly A RNA preparation
ELISAs
  Adenoviral vector construction and production
  Antigen ELISA or Titer (HBsAg) DiaSorin HBV Antibody to Surface Antigen ELISA or Titer (anti-HBsAg Ab)
  EBV: GenBio VCA IgM ELISA or Titer GenBio VCA IgG ELISA or Titer GenBio EBNA1 ELISA or Titer GenBio EA ELISA or Titer
  HBV: Ortho HBV Core Antibody ELISA or Titer (anti-HBVcore Ab) BioRad HBV Surface
  HCV: Ortho HCV ELISA or Titer In-house HCV peptide ELISAs
  HIV-1: Bio-Rad HIV-1 ELISA or Titer
  HTLV: Biomerieux HTLV-1/2 whole virus ELISA or Titer
  Lentiviral vector construction and production
Expression Optimization in E. coli
  Analysis for total and soluble protein and testing of purification and protease cleavage
  Custom BMG fluostar Optima plate reader software for testing multiple expression options in parallel
Fluorescence Anisotropy
  High through-put molecular screens
Gas Chromatography (GC)
  Determination of isotopically-labeled metabolic probes from cell extracts
  Quantitation of compounds-of-interest from human biofluids
Gene Cloning
  Completely sequenced entry clones
  Expression constructs transfected into expression hosts
  Over 150 Gateway expression vectors
  Subcloning of genes into expression vectors
High-Performance Liquid Chromatography (HPLC)
  Determination of modified DNA bases from cell extracts
  Metabolite profiling (metabolomics) using HPLC-MS
  Purity determination of proteins, peptides, and monoclonal antibodies
  Separation and quantitation of estrogen metabolites from human biofluids and tissue
  Separation of drug metabolites in human biofluids
Image Analysis
  Auto-location of non-straight dividing paths between touching cells and nuclei
  Deconvolution
  Enumerating individual FISH signals in cells within intact tissue
  Measurement of the spatial organization of cells within tissue
  Quantification of structural and molecular properties
  Quantifying average telomere length in populations of interphase nuclei
  Quantitative co-localization for measuring protein-protein interactions
  Segmentation of cells and subcellular components from specimen images
  Tiling of small images together into a single large-area image
Immunoelectron Microscopy (IEM)
  Negative-stained analysis of bacteria and virus samples
  Pre- and post-embedding analysis of tissues and cells
  Silver-enhanced analysis of tissues and cells
In-House Real-Time Quantitative PCR Assays
  EBV: EBV pol real-time Quantitative DNA-PCR (= EBV viral load)
  HBV: HBV x-gene real-time Quantitative DNA-PCR (= HBV viral load)
  HCV: HCV 5’UTR real-time Quantitative RT-PCR (= HCV viral load)
  HIV: HIV gag real-time Quantitative RT-PCR (= HIV viral load)
  HTLV: HTLV tax real-time Quantitative DNA-PCR (= HTLV proviral load)
  Mammalian Cell Quantitation: ERV-3 real-time Quantitative DNA-PCR
Instrumented Expression
  Six fully instrumented minifermenters that lower costs and increase quality
  Tank fermentations of E. coli, Pichia pastoris, and other microorganisms
Laboratory Automation
  Customized automation solutions
  DNA, RNA, and protein preparation
  Real-time PCR
  Sequencing
Light Microscopy (LM)
  Fluorescent, inverted, and stereo LM
  Semi-thin-sectioned macro analysis of tissues from resin-embedded block
LUMINEX Assays (for Cytokines, Chemokines and Cell Signaling Analytes)
  Sample types that can be assayed include: serum, plasma, cell lysates and culture supernatants.
MALDI-TOF Mass Spectrometry
  MALDI-TOF mass spectrometry studies of proteins, peptides, DNA, and carbohydrates
Mass Spectrometry (MS)
  Chemical ionization
  Electron impact ionization
  Matrix-assisted laser desorption (MALDI)
  Surface-enhanced laser desorption (SELDI)
Microarray
  CDNA; oligonucleotide; miRNA; virus
  High throughput gene expression
  Microarray CGH
mRNA Isolation, Purification and Characterization – Expertise
  Establishment of RNA isolation and purification protocols from tissues and cells from a variety of species
  Quality control analysis
  Real-time PCR
  RNA blot analysis
  Total or cytoplasmic RNA and poly A RNA preparation
Nuclear Magnetic Resonance (NMR)
  1D spectra with water/solvent suppression (1H, 13C, 15N, 19F, 31P)
  Consultation and data interpretation
  Engineering/hardware support
  Implementation of standard and custom pulse sequences
  Metabolite profiling (metabonomics) of human biofluids
  NMR and ESR electronic design improvements
  Structure determination of unknown compounds
  Through-bond correlation (COSY, TOCSY, HSQC, HMQC, HMBC, CIGAR-HMBC, 1,1-ADEQUATE)
  Through-space correction (TrROESY, NOESY)
Nucleic Acid Extraction, Purification & Analysis
  DNA Extraction - includes OD260/280 and ERV-3 real-time PCR (PBMC, lymphocytes, buffy coats, granulocyte fractions, tissues, etc)
  Robotic Liquid Handling & Sample Aliquoting
Protein Blots
  HCV: Ortho HCV RIBA (recombinant immunoblot assay)
  HTLV: MP Diagnostics HTLV Western Blot
Protein Identification
  Bioinformatic analysis of sequence data
  Chromatographic protein separation (ion-exchange, gel-filtration, reverse-phase chromatography, HPLC)
  One- and two-dimensional gel electrophoresis with preservation of biological activity
  Protein identification by Peptide Mass Fingerprinting (PMF) using enzymatic digestion and MALDI-TOF mass spectrometry
  Proteolytic chemical fragmentation and purification of resultant peptides and proteins
  Western blot analysis
Protein Purification
  Affinity purification and subsequent tag and protease removal
  Three computer-controlled chromatography stations at three volume capacities
Real-Time PCR
  DNA content
  mRNA expression
Scanning Electron Microscopy (SEM)
  Analysis of cell-surface immunolabeling
  Analysis of cultured cell and tissue samples
  Ultrastructural analysis of nanoparticles and elements using an EDAX-equipped SEM
shRNA Technology
  Consultation and shRNA Design
  shRNA Lentivirus production to generate lentiviral delivery systems for analysis of stable and functional gene silencing
  shRNA screening in which activity is determined through the use of cell-based assays including Beta-galactosidase, Renilla Luciferase and Firefly Luciferase assays
  Vector construction (in collaboration with PEL)
Supercritical Fluid Chromatography (SFC)
  Separation of complex estrogen metabolites and phyto-estrogens
Transmission Electron Microscopy (TEM)
  Analysis of bacteria, protein, nanoparticle, and virus samples
  Analysis of DNA, DNA-protein complexes
  Negative-stained and thin-sectioned TEM
  Semi-quantitative analysis of virus pellet samples
  Shadow-replica analysis of virus and virus core samples
  Ultrastructural analysis and viral diagnosis using 3D TEM
Health and Human Services (HHS) National Institutes of Health (NIH) National Cancer Institute (NCI) SAIC Frederick, Inc.