SY86 gene is one of the genes contained in the AZF region in the long arm of the Y chromosome that control male spermatogenesis. Previous research proved that this gene frequent deleted in infertile men. Purpose of this study is to make a construct sY 86 gene, which will be used to make antibodies to detect infertility in men. The material used cDNA fragment sY86 with molecular weight 326 bp derived from the blood of man with normal sperm category. This fragment was ligated in PET TOPO vector, and the ligation product was transformed to E.col TOP 10. Selection of transformants using the ampicillin because PET TOPO vector containing a gene that resistant to ampicillin. Plasmids then were isolated and the success of isolation showed by the result of electrophoresis. To ensure the size of the plasmid, the DNA fragment was performed by PCR method and PCR results were then followed elektrophoresis. if it showed appropriate band size (326 bp), the DNA was sequenced and the result that accordance with sY 86 gene from NCBI can be used for expression in liquid LB medium with antibiotics.
Keywords : cDNA, sY86, ligation, transformation, vector