Skills:
Basic molecular techniques: Western blot, Northern blot, Southern blot, electrophoretic mobility shift assays (EMSA), protein concentration determination (Bradford, Lowery, and TCA), spectrometry, fluorometry, confocal microscopy, fluorescent microscopy, protein staining (Ponceau Red, Ruby Red, Coomasee Blue), DNA concentration determination using PicoGreen, dual luciferase luminescence, quantification of apoptosis by Hoechst staining and caspase cleavage, polymerase chain reaction, oligo-nucleotide primer design, site directed mutagenesis, plasmid cloning, fluorescent DNA sequencing, nitric oxide analysis, enzyme-linked immunosorbent assay (ELISA), ELISA spot assays, multiplex cytokine analysis, liposome production of hydrophilic compounds in dilauryl phosphatidylcholine (DLPC) for inhalation delivery, dialysis, lyophilization, measurement of proteinase activity (ENZcheck and zymography), and production of allergens. Cellular techniques: DNA repair analysis (host cell reactivation, slot blot analysis using DNA-adduct specific antibodies, and strand specific repair southern blot), shuttle vector mutagenesis, Hprt mutagenesis, cell viability (MTT and AlamarBlue), colony forming ability, stable and transient transfection of eukaryotic cells (electroporation and phosplipid), siRNA in eukaryotic cells, differential cell analysis via cytospin and smear slides, isolation of immune cell populations (MACs and flow cytometry assisted cell sorting), isolation of peripheral blood mononuclear cells (PBMC), cell culture of primary and immortalized fibroblasts, EBV transformed lymphocytes, and spontaneously immortalized keratinocytes. nimal techniques (mouse and rat experience): Animal husbandry, housing, genotyping, breeding, organ isolation (blood, brain, lymph node, liver, lung, and spleen), organ homogenization for immune assays, inhalation anesthetic administration, intranasal challenge, tracheal intubation, intravenous cannulation/injection, intraperitoneal injection, gastric lavage, measurement of airway hyper-reactivity via whole body phythesmography, bronchial alveolar lavage (BAL) collection, lung preparation for histology, histological analysis (hematoxylin and eosin stain, periodic acid-Schiff stain, Grocott's silver fungal stain). Mycology techniques: Growth/isolation of fungi from environmental, animal, and clinical samples (BAL and sinus lavage), preparation of slides for fungal identification (lactophenol cotton blue and calcifor white staining), growth of fungi for allergen production, preparation of cryogenic conidia stocks for inhalation experiments, and fungal clearance and dissemination assays.