Marketable and relevant experience and expertise include:

MOLECULAR CLONING:

• Molecular cloning techniques including cloning scheme development for protein and gene expression and targeting vectors
• Backbone sourcing and modification
• Adapter primer design and polymerase chain reaction (PCR) condition optimization
• Restriction digestion
• Dephosphorylation via Calf Intestinal Phosphatase (CIP) and Shrimp Alkaline Phosphatase (SAP)
  
• Ligation via T4 Ligase
  
• Related bacteriology skills (below)

BACTERIOLOGY:

• Routine laboratory bacteriology to include production of competent cells
• Plasmid transformation via heat shock and electroporation
•  Plating and streaking for isolation
• Colony picking and blue-white selection
  
• Expansion in liquid culture including optical density monitoring
• Plasmid DNA purification via methods including mini-, midi-, and maxi-preps Cesium Chloride (CsCl) gradient banding, and crude purification via alkaline lysis.
  
• Procedures also include those relevant to protein expression and purification (below)

PROTEIN EXPRESSION & PURIFICATION:

• Transgenic protein expression induction using Isopropyl β- d-1-thiogalactopyranoside (IPTG)
• Bacterial cell lysis using lysozyme, benzonase, DNAse, RNAse, sonication, and other mechanical means
• Lysate separation via ultracentrifugation.
• Protein purification via conventional and fast protein liquid chromatography (FLPC, generally using GE ÄKTA)
• Protein quantification via Bradford assays
• Analysis of protein purity via Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) followed by Coomassie Brilliant Blue staining
• Buffer exchange via dialysis

CELL CULTURE:

• Routine and specialized cell culture techniques including isolation of primary skin fibroblasts
• Handling of a wide variety of primary and immortalized cells
• Splitting / subculturing adherent cells via typsinization
• Cell counting using both conventional hymocytometers and automated systems (e.g. the Countess II)
• Seeding / inoculation of plates and flasks
• Preparation and revival of cryopreserved cells.

ANIMAL HUSBANDRY / HANDLING / DISSECTION / TISSUE COLLECTION:

• Animal husbandry responsibilities to include matings, weanings, and euthansia
• Collection of blood, serum and plasma from live mice
• Blood gluclose monitoring in live mice, to include during insulin / IGF-1 tolerance tests
• Analysis of live-animal body composition via benchtop NMR (e.g. via Bruker MiniSpec)
• Development and performance of novel PCR based genotyping procedures for genetically modified mouse lines
• Dissection of a wide variety of mouse tissues
• Pulverization / homogenization of tissues for subsequent RNA and protein analyses

DRUG AND INTERVENTION TESTING/DEVELOPMENT:

• Conceive of, develop, and execute cell-culture experiments to examine drug kinetics, dynamics, interactions, serum-shift properties, and impacts upon biological pathways of interest via protein and RNA analyses (below).
  
• Execution of concurrent of subsequent analyses in animal models as appropriate.

PROTEIN ANALYSIS, WESTERN & IMMUNOBLOTTING:

• Preparation of lysates / protein extracts from cultured cells and pulverized/homogenized tissues
• Literature and market review of available and proprietary antibodies
• Production of dephosphorylated negative controls via treatment with lambda protein phosphatase (LPP/ λPP)
• Optimization of extraction ratios, protein loading, and primary and secondary antibody dilution levels during development of florescent (e.g. via LI-COR Odyssey CLx) and chemiluminescent western blots (e.g. via Fisher iBright FL1000), and capillary electrophoresis based immunoblots (e.g. via Protein Simple Wes)
• Quantification with ImageJ, ImageStudio, and other software, and subsequent data analysis.

RNA ANALYSIS, RT-qPCR, qRT-PCR:

• Purification of RNA from cultured cells and pulverized/homogenized tissues using Trizol and Qiagen RNeasy kits
• Quantification of RNA via NanoDrop or QuantIt RiboGreen assays
  
• Analysis of RNA integrity via Agilent 2100 bioanalyzer
• Generation of complimentary DNA (cDNA) via several commercial reverse transcriptase (RT) kits
• Generation of appropriate negative controls
  
• Analysis of assay limit of detection, linearity and amplification efficiency
  
• Measurement of cDNA abundance via SYBR Green and TaqMan / probe-based method (e.g. via an Applied Biosystems StepOnePlus) and subsequent data analysis

REGULATORY AFFAIRS:

• Experience working in accordance with Environmental Protection Agency (EPA) Good Laboratory Procedures (GLPs) and the Federal Insecticide Fungicide and Rodenticide Act (FIFRA)
• Experience working in accordance with Food and Drug Administration (FDA) GLPs including an ISO13485 compliant quality management system (QMS)
• Followed, reviewed, authored and amended Standard Operating Procedures (SOPs)
  
• Experience working in a Biosafety Level 2 (BSL2) laboratory
• Familiar with Health Insurance Privacy and Portability Act (HIPAA) requirements from work in university health systems
• Performed work in support of deregulation Cargill's Latitude Crop Product (produced in collaboration with BASF corp): https://www.cargill.com/page/latitude
  

VIROLOGY / BSL2:

• Isolation of unknown but pathogenic human viruses from patient clinical samples
• Amplification of viruses through cell culture to include examination of cytopathic effect (CPE)
  
• Serotyping and titration of viruses in fixed cells via immunostaining and light microscopy
  
• General procedures for safely working with such viruses in a Biosafety Level 2 (BSL2) laboratory

OTHER SKILLS:

Dr. Bower continues to develop a variety of skills for the benefits of both his clients and BDB LLC.
While these skills may not be at an expert level, they may serve as useful adjuncts for a client’s project.

• Software Development / Programming: Java, Python, SAS, C#
• Web Development: HTML, CSS, JavaScript, PHP, web server configuration
• Database Programming / Management: SQL, MySQL, Microsoft Access
• Real Estate: Property record research, property acquisition, holding and resale
• Finance: Creation of Expert Advisor (EA) systems on a hobby basis