Abstract Introduction/Objective Bacterial infection is a significant cause of morbidity and mortality. Prompt identification of microorganisms and their susceptibilities to antimicrobial therapies is critical in the management of patients with bloodstream infections. Blood cultures are collected in paired aerobic and anaerobic bottles. However, transport delays might allow some organisms to grow extensively prior to incubation in the blood culture instruments, leading to false-negative culture results. The Detroit Medical Center utilizes the BD Bactec™ instruments for blood culture incubation and the Verigene DNA-based molecular assay for the identification of bacteria and major resistance genes. It has a core microbiology lab that serves 6 hospitals, however, 2 of the hospitals are remotely located. The aim of this project was to determine if transportation delays led to significant false-negative culture results. If significant false negativity occurred, additional Bactec™ instruments would need to be purchased. Methods For one month, we tracked the collection of blood cultures to incubation time at one of the remote hospitals. All blood cultures that remain negative after 164 hours of incubation are routinely discarded. However, in this case, they were subcultured to a Petri plate containing chocolate agar for 30 days. Any organisms that grew were identified by standard lab techniques. Results Of the 547 negative culture bottles that were subcultured for possible false-negative results, only 3 (0.5%) bottles grew bacteria. All three were isolated from different patients. The mean time from blood collection to incubation in the instrument was 4-8 hours. The isolates either met criteria for contaminated cultures, or they grew the same pathogen that had previously been identified in the paired bottle from the same culture. The organisms isolated include coagulase negative Staphylococcus species, Staphylococcus pettenkoferi, and Pseudomonas aeruginosa. No unexpected pathogenic organisms were detected. Conclusion Our results demonstrate that prolonged pre-analytical time does not lead to false-negative blood culture results. The patients’ diagnoses were not changed, therefore, the purchase of additional blood culture instruments was not necessary. However, transportation time from the patient floors to the main microbiology lab needs to be improved to meet the recommended 2 hours pre-analytical time.