A one-step efficient and specific non-radioactive non-fluorescent method for in situ hybridization of banded...

A non-radioactive method for in situ hybridization of cosmid probes to metaphase chromosomes is described. Two procedures are involved: (i) hybridization with a cosmid probe labelled by nick translation in the presence of digoxigenin dUTP. The signal is visualized by an alkaline phosphatase conjugated antibody. (ii) FPG banding of the chromosomes. The steps involved in these two procedures are combined in an order which allows simultaneous observation of the banding pattern and the hybridization signal. The metaphases can thus be analysed after a single photographic step. This technique is considerably simpler than the method used previously.