Sex determination in animal by Multiplex Polymorase Chain Reaction

Abstract

The objective of this study was to develop a multiplex polymerase chain reaction (multiplex PCR) technique for sex determination of cattle and buffalo. Twenty-seven blood samples were collected from both sex of crossbred Holstein Friesian, native cattle, Brahman, and buffalo. DNA samples were extracted, and sex of animals were determined by multiplex PCR method. Two pairs of specific primers derived from the sequence of a SRY gene served as male-specific primers. They were used for a verification of Y chromosome. The Kappa-casein gene was used as an internal control. The results showed that the SRY gene specific primer was able to determine the sex of cattle and buffalo with high efficiency. The Kappa-casein gene primer was designed as an internal control for the cattle but not for buffalo.