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Equipment

The Khvorova Laboratory occupies approximately 6,000 square feet (bays AS4-1013 through AS4-1016) on the fourth floor of the new Albert Sherman Center at the University of Massachusetts Medical School (UMMS) Worcester campus.

Mid-Scale RNA Synthesis, Purification, and Quality Control System: With the support of a recent NIH S10 Shared Instrumentation grant (Khvorova, PI), UMMS has purchased a GE ÄKTA Oligopilot 100 system, an Agilent preparative HPLC, and an Agilent QTOF LCMS system. The instrumentation enables cost-effective mid-scale custom oligonucleotide synthesis, purification, and quality control, which are functions essential to generating compounds necessary to run in vivo studies.

RNA Synthesis: The Khvorova Laboratory is furnished with standard chemistry synthesis equipment (four rotovaps, twelve chemical hoods), five SpeedVacs, a Freezone PL2 lyophilizer, an ABI peptide synthesizer, an ABI Expedite 8909 oligonucleotide synthesizer, and a MerMade 12 oligonucleotide synthesizer.

RNA Analysis and Purification: The Khvorova Laboratory is equipped with four HPLC systems (1100 and 1200, with an auto-sampler, fraction collector, oven WVL detector, and fluorescence detectors), an Agilent 2100 electrophoresis bioanalyzer system, a Nanosight, a GE ÄKTA purifier system, Teledyne Combiflash chromatography, and Finnegan high resolution LC-MS (up to 2000 m/s). The lab also has standard biochemical equipment for running native PAGE gel electrophoresis for RNA analysis and uses a shared GE Healthcare LAS4000 imager for gel imaging.

Cell Culture: A fully equipped tissue culture room is located adjacent to the Khvorova Laboratory. It houses two 6' BSL2 tissue culture hoods, six CO2 incubators, a Nikon inverted fluorescence microscope, two dissecting microscopes, a 37°C water bath, a tabletop centrifuge, an automated cell counter, and other small equipment required for mammalian cell culture. Liquid nitrogen and -80°C freezer storage is available for all cell lines. This space also is used for purification of primary cell culture and tissue dissection.

Gene Expression Analysis: mRNA expression is analyzed by the QuantiGene 2.0 branched DNA assay. The Khvorova Laboratory houses two QuantiGene ovens, two plate washers, a Tecan Infinite M1000 Pro microplate reader (for fluorescence, luminescence, and absorbance), a Qiagen TissueLyser II, and all reagents necessary for automated gene expression analysis in 96 well format. A real-time qPCR machine also is available.

Histology: An alcove in the Khvorova Laboratory is dedicated to histological work and is equipped with materials, including a M900 easydip slide staining system to stain brain sections with DAPI and other antibodies.

Microscopy: Confocal microscopy is performed on a shared CSU10B Spinning Disk Confocal System scan head (Solamere Technology Group) mounted on a TE-2000E2 inverted microscope (Nikon) with a 60x Plan/APO oil lens and a Coolsnap HQ2 camera (Roper Scientific).

Additional Equipment: The Khvorova Laboratory has access to the equipment and resources of the RNA Therapeutics Institute and the UMass Research Cores.

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