To investigate pediatric low-grade gliomas for alterations in . DNA and RNA were extracted from 62 pediatric gliomas. Molecular methods included PCR, RT-PCR, and RNA sequencing; Sanger sequencing was used for validation. Analysis for hotspot genetic alterations in R132 and R172 (45 and 33 samples) was negative in all cases. deletions were detected in exons 1 and 2 in 1 (pleomorphic xanthoastrocytoma) sample of 9 samples analyzed. Of 10 samples analyzed for translocation, 4 each were positive for translocations with exon 2 and exon 3 of . Six samples showed rearrangement. The lack of genetic alterations is in accordance with the literature in pediatric tumors. Alterations in were recently reported to characterize diffuse grade II, but not grade I, gliomas. We optimized methods for analyzing gene variations and correlated the findings to pathological grade. The high incidence of MYB and MYBL need further evaluation. We also compared DNA, RNA, and RNA sequencing results for fusion, translocation, and genetic alterations. More accurate identification of the underlying biology of pediatric gliomas has implications for the development of targeted treatment.