Altered expression of oncogenes and tumor suppressor genes, the normal function of which is to regulate cell growth and differentiation, represents a central event in the pathogenesis of human cancer. Aberrant expression of these genes is often a result of a mutational event. In vitro amplification of DNA with the polymerase chain reaction (PCR) has enormously increased the sensitivity of the methods to detect mutations. These PCR-based techniques have thus become invaluable in the elucidation of the mechanisms of carcinogenesis as well as in molecular genetics. In addition, the precise definition of a mutation at the molecular level can be a very valuable adjunct to the diagnosis and classification of malignancies as well as to their prognostic assessment. In this article several PCR-based strategies are outlined, their applicability in the detection of different types of mutations is discussed, and finally the application of these techniques in fresh and archival tissues is presented.