Background: The VEGFA gene encodes vascular endothelial growth factor A (VEGF-A), which plays a key role in vasculogenesis and angiogenesis. The VEGFA rs3025039 (+936C>T) polymorphism is associated with many diseases. This study aimed to: (1) Apply PCR-RFLP technique to identify the VEGFA rs3025039 (+936C>T) polymorphism; (2) Primarily evaluate the distribution of genotypes and allele frequencies of the rs3025039 polymorphism in volunteers. Materials and methods: DNA extraction was isolated from peripheral blood of 60 volunteers. Determining the VEGFA rs3025039 (+936C>T) polymorphism by PCR-RFLP technique. Confirming the results of the genotypes randomly by Sanger sequencing. Results: All PCR-RFLP results of validated samples were in concordance with sequencing results. The distribution of CC, CT and TT genotypes by rs3025039 polymorphism accounted for 80%, 16.7% and 3.3%, respectively. The frequencies of C and T alleles were 88.3% and 11.7%, respectively. Conclusion: Successfully applying PCR-RFLP technique to determine the VEGFA rs3025039 (+936C>T) polymorphism, which establishes the groundwork for further research into the association between this polymorphism and various disorders.