Production of bovine Y painting probes through chromosome microdissection and DOP-PCR

We are successfully suceeded in producing 2 painting probes from the bovine Y chromosome using chromosome microdissection and DOP-PCR techniques. We are separately microdissected 5 to 6 copies from both the whole Y bovine chromosome and the q arm of the Y bovine chromosome. We amplify these microdissected DNA fragments through DOP-PCR. Fluorescent in situ hybridization technique (FISH) with these probes on a metaphase spread from a normal bovine lymophcyte cultures from cattle, buffalo, sheep and goat was applied. Surprisingly that both of the two probes were hybridized only to the bovine Y chromosome without any background or cross hybridization to any other chromosome especially the X chromosome. A similar hybridization signal was also observed when these probes were tested in river buffalo. In contrast, no signal was observed when these probes were tested on both sheep and goat metaphase spreads.