Pathogen contamination and harborage in low-moisture food (LMF) processing environments have resulted in outbreaks and recalls, but researchers are limited in their abilities to investigate solutions. Methods used in most laboratory studies do not accurately reflect the route of contamination or harborage of pathogens in LMF environments, which complicates studying of sanitation methods. Inoculation methods were compared to establish low-moisture food persistent bacterial populations (LMF PBPs) that realistically reflect populations found in LMF environments. Culture-preparation techniques, food matrices, and ratios of nutrient to moisture were compared for their impacts on Salmonella Tennessee survival. A slurry method with 0.5 mL of culture to 0.4 g (NFDM) was easiest to form, inoculate, and process for further investigation, as was more consistent than other methods. With the chosen technique, LMF PBPs of Salmonella Tennessee (0.5: 0.4 [mL Salmonella Tennessee to g Powder]) with different powders (NFDM, peanut butter powder [PB], chicken powder [CP] and silicon dioxide [SiO