Detection of telomerase activity utilizing energy transfer primers: comparison with gel- and ELISA-based detection.
We have developed a closed-tube format telomeric repeat amplification protocol (TRAP) assay for direct quantification of telomerase activity within the PCR vessel. The assay utilizes energy transfer (ET) primers, which emit fluorescence only upon incorporation into PCR products. This novel ET primer system (Amplifluor primers) has major advantages over existing detection methods because it eliminates the need for post-PCR processing and thus reduces greatly the risk of carryover contamination and the time required for the sample analysis. The assay is as sensitive, specific and quantitative as the polyacrylamide gel-based or ELISA-based TRAP assay.
Uehara
H
H
Intergen Company, Gaithersburg, MD, USA.
Nardone
G
G
Nazarenko
I
I
Hohman
R J
RJ
D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005452:Fluoresceins; D005453:Fluorescence; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D012680:Sensitivity and Specificity; D019098:Telomerase / Q000032:analysis* / Q000378:metabolism; D016615:Telomere / Q000201:enzymology / Q000235:genetics
D017931:DNA Primers; D005452:Fluoresceins; D019098:Telomerase