MOLECULAR-BASIS OF ARTIFACTS IN THE DETECTION OF TELOMERASE ACTIVITY AND A MODIFIED PRIMER FOR A MORE ROBUST TRAP ASSAY

Human somatic cells have essentially no telomerase activity, Telomeras e is linked to tumor genesis and is a valuable marker far malignant gr owth, Extreme paucity of the enzyme neccessitated development of a PCR -based assay, 'telomeric repeat amplification protocol' (TRAP), Unfort unately, this method is not without difficulties, Amplification produc ts are not related to the size of the amplified telomerase products, F urthermore, false positive results can occur, and careful control of r eaction conditions is crucial. We analyzed in detail the molecular bas is of artifacts, Based on these data, reverse PCR primer was changed a nd both problems in the TRAP assay were eliminated.