Grant: Research Tools and Instruments - Category 1 (<$150,000)

The goal of this application is to acquire a specialized cell disrupting and ultracentrifugation system for the preparation of protein extracts, isolation of organelles, and fine separation of molecular complexes. Cell lysis is the first step in cell fractionation, and as such, opens the door to a myriad of biological studies. All cells have a plasma membrane, a protein-lipid bilayer that forms a barrier separating cell contents from the extracellular environment. For bacteria (especially Gram-positive), fungi, and plant cells, the plasma membrane is also surrounded by a cell wall, which is a composite structure that consists of polysaccharides and proteins. The cell wall is both firm-to-rupture and elastic, thereby fortifying the cell from hostile environments encountered by the organism. Because our studies use bacteria, yeasts, and plant cells as model systems, it is crucial that we can have access to a highly efficient multi-sample bead cell disrupter in order to efficiently extract DNA/RNA, protein, and organelle content from resistant biological samples. Once whole cell extracts are prepared, our research groups subsequently require the use of an ultracentrifuge to perform differential pelleting of organelles, microsomes, ribosomes, and proteins. Figuring prominently in the purification protocols, ultracentrifuge relies on the same physical principle that makes children dizzy on playground carousels. As the rotor spins, objects are pushed away from its axis of rotation via centrifugal force. The ultracentrifugation force could approach more than 1,000,000 times that of gravity, thereby allowing differential pelleting of cellular components as their size/weight decrease. In this instrument grant application, we opted for a tabletop model of ultracentrifuge because smaller volumes of samples can be spun. Furthermore, it offers higher speed, which allows faster separation of cellular components. With an efficient micro-ultracentrifuge, we will reduce separation time, thereby diminishing overall turnaround time, as well as improving work flow and productivity.