ABSTRACT
There are several critical steps required to obtain a chromosome-specifi c probe for fl uorescence in situ hybridization (FISH). Initial DNA is obtained by mechanically microdissecting selected regions of target chromosome in metaphase spreads, followed by an unspecifi c Polymerase Chain Reaction (PCR) amplifi cation and a subsequent regular stringency PCR to obtain quantitative amplifi cation of the microdissected DNA. Finally, the DNA must be labelled in order to function as a FISH probe. Every step can be individually checked but only once the procedure is fi nished, in other words when the hybridization is developed on the slide, we can ascertain a successful procedure.
