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Twin marker genes are the controlling genes of twinning which can increase the frequency of twin ovulations in one estrus cycle. The role and function of genes in determining twin births is very important to understand in order to consider the potential of females that can produce twin. Detection and identification of twin controlling genes taken from twin cows, where sires give birth to twins or twin tillers. The aim of this study was to identify sequences of twin marker genes in cattle and analyze the function of twin cow controlling genes (Bos taurus) from the results of sequence variations. This research is included in exploration research which has procedures including DNA isolation, PCR, electrophoresis analysis, and bioinformatic analysis. The results of this study were detected by the twin controlling gene with a birth weight gene marker in the six cow blood samples (Bos taurus) at 56oC attachment temperature. The results of the analysis of the diversity of nucleotide sequences from the six blood samples showed various variations and the analysis of the results of protein expression that could be identified from the birth weight gene marker was alpha V beta 3 integrin (αvβ3).