Genetic Engineering and Viral Vector Core – Equipment

The GEVVC provides training and instruction materials for the safe and responsible use of relevant capital equipment including shaking incubators, centrifuges, ultracentrifuges, gel documentation system, capillary electrophoresis systems, droplet digital PCR system, and protein purification system.  Due to the biosafety considerations involved with viral vectors, access is granted on an as-needed basis once appropriate training has been given.  Equipment marked with (*) are held under MMCSI but are shared via GEVVC.

Agilent (AATI) 5200 Fragment Analyzer is an automated capillary electrophoresis system that is capable of high sensitivity and high-resolution characterization and quantification of nucleic acids. It is used in our standard QA assays during plasmid and virus production as well as conventional and CRISPR-related genotyping assays.

Azure C200 gel documentation system is capable of the UV and visible fluorescence imaging after gel electrophoresis. This device is located within the BSL2 virus production lab.

Azure C600 gel documentation system is capable of imaging UV, visible, and infrared fluorescence of samples after electrophoresis or after transfer to membrane substrate and exposed to labeled probes. It is also capable of recording chemiluminescent signal for high-sensitivity analysis.

Beckman Coulter Avanti J-26 XPI centrifuge is used for harvesting bacteria and/or mammalian cells grown in culture, as well as during the isolation nucleic acids and proteins from cell lysates. A variety of rotors are available to accommodate different tube and bottle volumes.

Beckman Coulter Optima L-100 XP ultracentrifuge is used for the isolation of organelles or viral particles from cell lysates.  A variety of rotor sizes are available, including swinging bucket and fixed angle options.

*Bio-Rad QX200 droplet digital PCR is used for the detection and absolute quantification of specific nucleic acids from DNA and RNA samples. It is used to establish the titer (viral genomes per mL) of our virus preparations and for the determination of copy number for our transgenic animal and cell-line models.

Denovix CellDrop is a microvolume DirectPipette™ cell counter capable of quantifying concentrations of cells in suspension, and assaying for transfection efficiency (% GFP-positive) or viability (trypan blue or acridine orange/propidium iodide).

Denovix DS-11 FX is a microvolume spectrophotometer/fluorometer used for quantifying nucleic acids (dsDNA, ssDNA, RNA) or proteins in a liquid sample.

GE ÄKTA FPLC is used for the automated affinity chromatography of adeno-associated virus and recombinant proteins.

New Brunswick Scientific I2500 shaking incubators are used for growing bacteria during plasmid and protein production at 32C and 37C.  Brackets, racks, and clamps for 14mL tubes and various sizes of Erlenmeyer flasks are available (50mL – 2L).

Oxford Nanopore MinION Mk1C is a “third generation sequencing device” for long-read/single molecule analysis of nucleic acids.  It is capable of sequencing DNA from various sources (plasmids, viral vectors, cells).  It is also capable of sequencing cDNA and RNA samples.

Coming Online Soon (TBA, post-pandemic):

Beckman Coulter Biomek 4000 is an automated liquid handler for ensuring precision and accuracy in standardized procedures for nucleic acid and protein isolation and purification.  This machine uses Solid Phase Reversible Immobilization (SPRI) Methodology for the automated purification of high molecular weight genomic DNA from cell pellets and biopsies for long-read sequencing applications.