ONCOGENE AMPLIFICATION SCREENING BY LABELED PRIMER MULTIPLEX POLYMERASE CHAIN-REACTION

This paper describes an improved procedure for rapid detection of ampl ified genes in fresh or formalin-fixed, paraffin-embedded tissues. Uti lizing a multiplex differential polymerase chain reaction with radioac tively labeled primers and electrophoresis of the products through thi n gels, it is possible to screen for oncogene amplification more rapid ly and reproducibly than has been previously demonstrated. This proced ure takes advantage of thin vertical gels with external cooling, which allows sharp band resolution. Four separate gels can be electrophores ed at the same time in a single gel box. Because each gel slab contain s 10 or more lanes, 40 or more samples can be assayed for gene amplifi cation simultaneously. The entire procedure can be carried out from fo rmalin-fixed, paraffin-embedded tissue to finish in 8 h when combined with a sonication technique for DNA extraction.