The growth differentiation factor 9 (GDF9) gene (exon 1) is the subject of this study's investigation into polymorphisms concerning fertility in Egyptian Baladi goats. Fifteen females that produce single kid and 55 produce twins kids were used in this study. Methods: blood samples were taken, and genomic DNA was extracted. By using specific primers, a 770 bp region of the GDF9 gene was amplified; the amplified products were subjected to sequence analysis and examined to determine the phylogenetic relationship. The sequence of GDF9 gene of Baladi goats was compared with eight Capra hircus accession numbers found in Gen Bank. The results of sequence comparison indicated that the Baladi goat is closely related to the breed of accession number KY780296. In addition, polymorphisms were covered by the PCR-RFLP technique using the Msp1 restriction enzyme to confirm twinning molecular markers. The banding patterns of Msp1 enzyme digestion showed three bands of approximately 770, 470 and 300 bp, for does producing twins and one band of approximately 770 bp for does producing single kid. Knowledge of the mutation in the GDF9 gene (exon 1) early on can be used to create a flock that specializes in producing twins without waiting for maturity and thus reducing production costs and increasing profitability.