Laboratory. Dr. Thrash’s lab (1,500 square feet) is located in the Allan Hancock Foundation building at the University of Southern California. The lab contains all the wet lab equipment needed to run the project. The laboratory is equipped for large scale water collection and transport with 1-20 L polycarbonate carboys, filtration equipment (Cole-Parmer Masterflex I/P peristaltic pump; 142 mm filter sets, 55 mm, 47 mm and 25 mm; Millipore Pellicon cross-flow filtration set; tubing), a -20˚C and -80˚C freezer, a refrigerator, 6 dry incubators, portable cold incubators, complete molecular biology facilities for the nucleic acid extraction and PCR (bead-beater, vortex, dry baths, water baths, thermal cyclers and BioRad GelDoc EZ Imager, gel trays and power supplies, Life Technologies Qubit fluorometer, refrigerated and unrefrigerated Eppendorf microcentrifuges and benchtop hanging bucket centrifuge), AirClean 24″ portable laminar flow workstation, portable cold incubators, optode PreSens system, vacuum pump, fume hood, Millipore Milli-Q water purification system, a full suite of chemical stocks, scales and a pH meter. The lab also has an OpenROV.
Culture infrastructure. The Thrash Lab has a dedicated culture room containing a Millipore Guava EasyCyte 5HT HPL benchtop flow cytometer for counting cells using a 488 nm laser (which can be transported and deployed on board), a Class II/A biosafety cabinet for non-confinement media inoculation and culture manipulation, an Eppendorf epMotion 5075 fluid handling robot with HEPA filtered chamber, and a BD Influx fluorescence-activated cell sorter with lasers from 355nm, 488nm and 640nm. Elsewhere in the lab is a ThermoScientific Locator 6 Plus liquid nitrogen dewar for storage of culture collections, and a dedicated benchtop and reservoir storage system for anaerobic culture. We also have a COY class A anaerobic chamber and a microaerophilic glove box. We have a full suite of culture vessels, including serum bottles, 50 custom 96 x 2.1mL PTFE plates, and polycarbonate and PTFE bottles of various sizes. Finally, the laboratory has an acid bath for a clean culture of traces of metals.