The polymerase chain reaction (PCR) is a rapid and highly sensitive method for detection of a variety of chromosomal translocations in malignant tissues. Detection of each different type of translocation, or even DNA rearrangements at different breakpoint cluster regions within the same type of translocation, usually requires separate thermocycling parameters and/or buffer conditions. In this report, we describe a single set of reaction conditions, making use of progressively decreasing annealing temperatures and a standardized reaction buffer, that permits the detection of several different translocations simultaneously. Specificity equal to or better than current procedures and sensitivity equivalent to one malignant cell in 1 |MX 105normal cells was achieved for translocations t(14;18)(q32; q21), t(9; 22)(q34: q 11), and t(4; l1)(q21; q23). For PCRs formerly requiring different, fixed annealing temperatures, the new technology allows batching or multiplexing of PCR samples. Thus, shorter turnaround time, decreased cost per sample, and simplified mechanization of PCR may be attainable using this assay.