Characterization of Chromosomal Rearrangements Using Multicolor-Banding (MCB/m-band)

Molecular cytogenetics and especially fluorescence in situ hybridization (FISH) banding approaches are nowadays standard for the exact characterization of simple, complex, and cryptic chromosomal aberrations within the human genome. FISH-banding techniques are any kind of FISH techniques, which provide the possibility to characterize simultaneously several chromosomal subregions smaller than a chromosome arm. FISH banding methods fitting that definition may have quite different characteristics, but share the ability to produce a DNA-specific chromosomal banding. While the standard techniques such as G-bands by Trypsin using Giemsa banding lead to a protein-related black and white banding pattern, FISH-banding techniques are DNA-specific, more colorful, and thus, more informative. At present, the most frequently applied FISH banding technique is the multicolor banding (MCB/m-band) approach. MCB/m-band is based on region-specific microdissection libraries, producing changing fluorescence intensity ratios along the chromosomes. Here we describe the FISH-banding technique MCB/m-band and illustrate how to apply it for characterization of chromosomal breakpoints with a minimal number of FISH experiments.