The need for bilateral pediatric bone marrow biopsies for cytogenetic and fluorescent in situ hybridization (FISH) studies has not been clearly delineated. We retrospectively identified 166 pediatric bilateral bone marrow biopsy specimens obtained from patients with a variety of clinical diagnoses, including solid tumors, lymphoma, leukemia, and other hematologic conditions. The cases included all pediatric bilateral bone marrow biopsies performed at our hospital spanning the years of 1992 to 2008. Agreement of FISH and classical cytogenetic results between the 2 sides was assessed. Of a total of 166 bilateral cases, 2 cases showed disagreement (1.2%), both from patients with solid tumors. One case was a rhabdomyosarcoma, in which FISH only was performed; the second was a neuroblastoma in which FISH and cytogenetics were performed (both FISH and classical cytogenetic results disagreed). The remainder of the cases showed complete agreement between the 2 sides (total 98.8%). We conclude that it is usually not necessary to perform bilateral bone marrow biopsies for FISH and cytogenetics in the pediatric population outside of the setting of solid tumor staging.