How do label DNA probes for whole chromosme paints and how do label for Spectral karyotyping?
Multicolor karyotyping labels and identifies each human chromosome pair by a different fluorescent color, using combinatorial labeling. Two groups using two different experimental strategies, have pioneered the procedure using five or six different fluors (SKYand M-FISH, respectively). Several M-FISH labeling schemes can be found here. Another group has achieved color karyotyping (Color Changing Karyotyping = CCK) using only three fluorophores, without the need for any specialized equipment or software. In M-FISH, five fluorophores are used to acquire the desired 24 combinations. Individual chromosome painting libraries and many individual fluorophores were tested, either separately or in groups . DNA amount for the various painting probes were adjusted, to bring the signals to comparable intensities. DNA template for all chromosomes labeled with the same dye was mixed together, resulting in five different pools. Five separate labeling reactions (nick translation or PCR) were performed
