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A novel closed-tube format telomeric repeat amplification protocol specifically adapted to real-time detection and quantification of telomerase activity was developed. The assay utilizes energy transfer primers, which emit fluorescence only upon incorporation into polymerase chain reaction (
PCR
) amplification products. The assay, performed on a real-time detection instrument, is highly reproducible, sensitive, and specific. Telomerase activity in as few as 10 cultured cells can be quantified with a linear dynamic range more than 2.5 logs. In addition, the presence of potential PCR inhibitor(s) is readily detectable by inclusion of an internal PCR control labeled with a second color fluorescence.