A simplified protocol for fluorescence in situ hybridization with repetitive DNA probes and its use in clinical cytogenetics

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Abstract
A method for chromosome-specific staining and its use in clinical cytogenetics is described. This fluorescence in situ hybridization protocol for repetitive DNA probes results in yellow-green fluorescent signals on orange-red stained chromosomes. Special characteristics are its simplicity, the use of digoxigenin-11-dUTP for labeling, and the combination of high stringency criteria for hybridization and low stringency for washing. The method is particularly advantageous in cases with structurally abnormal extra chromosomes (ESACs), marker chromosomes of gonosomal origin, and chromosomal mosaicism. It may also facilitate the screening of cases for fragile X. The chromosome-specific staining can be done within 1 working-day.