PCR conditions
PCRs consisted of 25 µl total volume with 5 ng purified BAC DNA, 0.2 µM primers, 0.2 mM of each dNTP, 2M MgCl2, 0.5 U Taq polymerase and 1 x PCR buffer. Thermal cycling included a 2 min denaturation step at 94°C, 30 cycles of 20 s at 94°C, 20 s at 58°C, 20 s at 72°C, and final extension at 72°C for 5 min. Analysis of PCR products was performed by electrophoresis on 1.8% agarose gels, and visualized with ethidium bromide staining.