Abstract

In this paper, we report identification of a novel male-specific DNA marker in Iranian river buffalo presenting the possibility of river buffalo gender determination using Random amplified polymorphic DNA-PCR (RAPD-PCR). Twenty eight random primers used for RAPD fingerprinting in order to isolation and characterization of microsatellite sequences using PIMA (PCR-based isolation of microsatellite array). All sharp bands related to different primers were isolated from the gel and inserted to a cloning vector and transformed to competent cells for nucleotide sequencing. Sequences were used as queries for BLAST tool. BLAST results showed that a sequence had high similarity with a part sequence of cattle Y chromosome sequences deposited in GenBank. A pair of primers (IRBMS-F and -R) was designed based on the sequence for amplifying the male-specific band by polymerase chain reaction (PCR) for Iranian river buffalo gender detection. Gender-specific bands in the gel were observed in all males but not in females. This showed that the gender of Iranian river buffalo could be easily and effectively identified by PCR technique using these primers.

Key words: Random amplified polymorphic DNA-PCR, gender detection, river buffaloes, male specific DNA marker.