Avidin fluorescein green

Mitotic chromosome preparation and FISH procedures were conducted using a modified protocol (Wang et al. 2001b ). Biotin-labeled and digoxigenin-labeled probes were detected by avidin-fluorescein (green) and anti-digoxigenin-rhodamine (red) (Roche Diagnostics, USA), respectively. Chromosomes were counterstained by

4’,6-diamidino-2-phenylindole

(DAPI) in antifade VECTASHIELD solutions (Vector Laboratories, Burlingame, CA). The concentration of block DNA (genomic DNA) was 200 times that of the chromosome-specific BAC DNA. The hybridization signals were observed using a fluorescence microscope (Leica MRA2) with a

charge-coupled device

(CCD) camera. Final image adjustments were performed using Adobe Photoshop CS3 software.

Mitotic chromosomes preparation and FISH procedure were conducted using a modified protocol [61 ]. Biotin-labeled and digoxigenin-labeled probes were detected by avidin-fluorescein (green) and anti-digoxigenin-rhodamine (red) (Roche Diagnostics, USA), respectively. Chromosomes were counterstained by 4′,6-diamidino-2-phenylindole (DAPI) in the antifade VECTASHIELD solutions (Vector Laboratories, Burlingame, CA). For the probe-cocktail mixture, gDNA was used as block DNA. The dose of block DNA was 200 times of the chromosome-specific BAC DNA. The hybridization signals were observed using a fluorescence microscope (Leica MRA2) with a charge-coupled device (CCD) camera (Zeiss Axioskop2 plus). Final image adjustments were performed by using Adobe Photoshop CS3 software.