The polymorphism CYP1A2*1F of the cytochrome 1A2 gene affects the inducibility of this cytochrome, which is involved in the metabolism of many commonly used drugs. We developed a rapid-cycle polymerase chain reaction (PCR) and fluorescent probe melting curve analysis on the LightCycler instrument for identification of this mutation. This new method was confirmed by genotyping 101 samples in parallel to restriction enzyme digestion. The 100% concordance of both methods showed the reliability of our high-speed genotyping assay, which is suitable for large epidemiological studies or routine clinical use.