A FLUORESCENT METHOD FOR DETECTION OF TELOMERASE ACTIVITY

The telomere repeat amplification protocol (TRAP) was recently develop ed to detect telomerase activity in cellular protein extracts. Teleome rase synthesizes a specific repeating nucleotide sequence onto the end s of telomeres, which stabilize eukaryotic chromosomes. Telomeric repe ats are identified in the TRAP method by polymerase chain reaction amp lification and incorporation of radionucleotides detected by autoradio graphy. Several drawbacks to this method have been recognized, includi ng the time required to complete the assay, the resolution of the resu lts, and the hazards of radioactive material. We have developed a new fluorescent method of detecting telomerase to alleviate these problems . Telomeric repeats are identified in the fluorescent TRAP (F-TRAP) as say by incorporation of fluorescein-labeled primers during amplificati on and subsequent detection with an automated DNA sequencer. This new method appears to be as sensitive as the standard TRAP assay and offer s advantages in speed, resolution, cost, and safety.