There is a common belief and practice that any exposure to oral or parenteral antibiotics prior to bone biopsy makes culture results unreliable. The aim of this article was to evaluate the effect of antibiotic exposure on bacterial yield in DFO microbiology specimens. The authors retrospectively evaluated 114 patients with DFO confirmed by histology. The primary outcome measurement was the proportion of bone biopsies with positive bacterial cultures. There was no statistically significant difference in culture yield in patients who received antibiotics (77.9%) and patients who did not (85.7%, P = .58). This study demonstrates that there were no differences in bacterial yield whether antibiotics were withheld or administered before bone cultures were obtained. The duration of antibiotic use prior to bone biopsy did not change the bacterial yield.