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MAJOR EQUIPMENT Specific Fields Relevant for Mouse Core Users The Emory Mouse Transgenic and Gene Targeting Core (Mouse Core). The Mouse Core is located on within the virus and antibody free (VAF), specific pathogen gree (SPF) vivarium in the newly opened, state of the art Health Sciences Research Building. The core consists of dedicated wetlab space and office space within the mouse facility as well as an office that is accessible for users. The Mouse Core’s laboratory areas include tissue culture space, procedure rooms, and an animal room dedicated to the Core. The Mouse Core infrastructure for core services include: Transgenic Mouse Production: • Diaphot 200 Nikon Inverted Microscope, Narishige MM-89 Micromanipulators, Narishige MO-188 Micromanipulator controllers: The microscope, micromanipulators, and controllers combine to create a system for the microinjection of 1-cell mouse embryos with a plasmid-based or BAC-based vector solution. • Micropipette Puller - P87: For preparation of microinjection needles to the desired parameters. • Allegra 64R Centrifuge, Beckman Coulter: Used to remove particulate matter from vector solution. • Sanyo MCO-5AC CO2 Incubator. This unit maintains temperature and CO2 levels during injection. Knock-out/Knock-in Mouse Production: • Axiovert Inverted Microscope, Narishige MM-89 Micromanipulators, and controllers: The microscope and micromanipulators enable the injection of murine ES cells suspended in media into blastocyst stage embryos. • Gene Pulser II + Capacitance Extender Plus: Used to electroporate murine ES cells with a targeting vector to initiate homologous recombination. • NuAire IR CO2 Incubators (2 units): These units maintain temperature and CO2 levels for culturing ES cells. • NuAire Biological Safety Cabinet/Nu-425-400 and Forma Biological safety Cabinet/1184: Biosafety cabinets where ES cell work is performed. Cryopreservation/Rederivation: • Bi-Cool BC IV-40 . Controlled-rate freezer that precisely controls freezing rates to -40 degrees Celcius. • Thermolyne Locator 4 LN2 Storage Unit, MVE XC47/11 LN2 Storage Units (5 units). Used to store cryopreserved embryos and sperm. 3 units are located in the core and samples are in duplicate in other 2 units at a backup location on a distinct power grid. • Sanyo MCO-5AC CO2 Incubator. This unit maintains temperature and CO2 levels during the IVF. Other: • Non-instrument computers - There are currently five Dell PCs used by the staff of the Core. • Forma Scientific Cryoplus LN Storage Unit: This unit is used by the Core to store murine ES cells for ongoing and completed projects. The unit has a liquid-level alarm which monitors the level of the liquid nitrogen at all times and alarms when a set low-limit is reached. • Thermo IEC CL3 Multispeed Centrifuge.