Detection of sex chromosome-specific DNA sequences by polymerase chain reaction using hair DNA in cattle

To establish a simpler polymerase chain reaction (PCR) strategy for detecting sex chromosome-specific DNA sequences in cattle, we verified the feasibility of using hair-derived DNA as a PCR template. In the control steer, multiplex PCR using hair-derived DNA resulted in the amplification of two amplicons: one derived from the X chromosome-harboring 1.715 satellite DNA (365 bp) and another from the Y chromosome-harboring btDYZ-1 sequence (199 bp). In heterosexual twin females with XX/XY chimerism confirmed by chromosome analyses, multiplex PCR analyses with hair DNA determined both 365 and 199 bp amplicons in five of the six cases. In the remaining case, multiplex PCR analysis using hair DNA revealed the amplification of only the 365-bp product; however, using singleplex PCR, the 199-bp product was successfully amplified from hair DNA. The results of the present study suggest that sex chromosome-specific DNA sequences can be detected by PCR using bovine hair-derived DNA.