Dylan Dufek Email & Phone Number

United States

Carlsbad, California, United States

Designed and carried out various experiments for the isolation of extracellular vesicles via PEG precipitation, microfluidic immunocapture, size exclusion column purification. Performed downstream molecular analysis of extracellular vesicle samples including RNA isolation, cDNA synthesis, RT-PCR detection of mRNA markers, digital PCR detection of mRNA markers, ultra-low input RNA-sequencing library preparation, and nanoparticle tracking analysis.Identified relevant microRNA markers for both extracellular vesicle clinical and quality control relevance and detected these markers through microRNA cDNA synthesis and RT-PCR detection.Developed several SOPs for downstream molecular processing and analysis of isolated cell/EV samples.Constructed target cell identification algorithms using imagePRO analysis software to identify specific target cells expressing certain immunofluorescent stains within a background of non-target cells deposited onto slides or micropatterned sievewell slides.Utilized flow cytometric cell sorting (namocell) to collect individually targeted cancer cells and performed subsequent minimal cell input RNA-sequencing library preparation.Conducted fluorescence microscopy on various immunostained cell preparations and executed single cell cell picking of individual target cells using micromanipulation modules with both capillary vacuum aspiration and laser capture microdissection.Performed whole genome amplification and DNA library preparation on isolated single cells.Quantified and sized RNA/DNA isolations and library preparations via Qubit, Bioanalyzer, and Tapestation systems.

Isolated viral RNA from both human collected samples and heat inactivated SARS-CoV-2 viral particles and carried out multiplex RT-qPCR to assess viral loads of each sample.Assisted with the research and development of a microfluidic assay for the capture and release of SARS-CoV-2 viral particles to distinguish between infectious and non-infectious saliva samples. Optimized sample processing parameters for microfluidic chips on both Hamilton StarLET liquid handling platforms and traditional syringe pump systems.Carried out routine tissue culture maintenance for various cancer cell lines.Developed a QC assay to assess the manufacturing surface chemistry coverage of a specific molecule by utilizing fluorescent imaging of microfluidic chips, generation of fluorescent intensity data using ImageJ, and using a custom python script to extract and process the data for manufacturing quality control. Cataloged years of experimental control data to assess the performance of microfluidic chips across different variables such as storage age, storage temperature, and cell line used. Utilized multivariate linear regression to evaluate optimal parameters for increased performance.

Chemically surfaced microfluidic chips using EDC-NHS chemistry with specific DNA oligos and capture antibodies. Chemical surfacing was performed with Hamilton StarLET liquid handlers.Effectively reduced antibody consumption for microfluidic chip surfacing by 30% without any change in downstream performance of antibody capture.Designed and performed various storage and stability tests for both microfluidic chip storage and reagent plates. Assayed reagents over the course of months using a microplate reader to assess turbidity and growth of bacteria.Acquired working knowledge and proficiency with Hamilton venus programming language in order to develop custom liquid handling steps and processes.

Assembly and running of several protein characterization techniques including BCA assays and ECL western blotting to quantify and identify proteins.Assist researchers through the modeling of cancer growth by fission-yeast cultures. Assist with autoclaving, media preparations, buffer preparations, sample acquisition, and stocking and maintenance of tissue culture inventory/supplies.