Abstract: To further analyze the molecular characteristics and detect the specificity of the transgenic soybean E8A7027 transformant with AgGlpF gene, the proposed study isolated the flanking sequence of transgenic soybean event E8A7027 and established a specific PCR detection system. Based on genome resequencing and Sanger sequencing, the sequence information obtained by genome resequencing analysis was aligned with the reference soybean genome to determine the insertion loci, the flanking sequences at 5′ and 3′ ends of T-DNA region. The PCR primers were designed according to the event-specific sequence, and the specificity and sensitivity of the PCR assay were vertified in the laboratory. The results confirmed that the integration site of transgenic soybean E8A7027 was Chr09: 33886331 with a single-copy insertion. The recipient genome DNA missed a 58 bp sequence at the insertion loci. The 910 and 802 bp flanking sequences at 5′ and 3′ end of E8A7027 event were obtained, respectively. The event-specific PCR with primers designed based on the two flanking sequences was enable to specifically distinguish the GM soybean event E8A7027 from other GM crops and non-GM soybeans, the sensitivity of this assay reached 0.05%（w/w）. The result can provide technical support for the safety evaluation and regulatory of E8A7027 transformants.