Nonradioactive detection of telomerase activity using the telomeric repeat amplification protocol.
The telomeric repeat amplification protocol (TRAP) is a two-step process for analyzing telomerase activity in cell or tissue extracts. Recent modifications of this sensitive assay include elimination of radioactivity by using a fluorescently labeled primer instead of a radiolabeled primer. In addition, the TRAP assay has been modified for real-time, quantitative PCR analysis. Here, we describe cost-effective procedures for detection of telomerase activity using a fluorescent-based assay as well as by using real-time PCR. These modified TRAP assays can be accomplished within 4 h (from lysis of samples to analysis of telomerase products).
Herbert
Brittney-Shea
BS
Department of Medical and Molecular Genetics, Indiana University Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana 46202-5251, USA. brherber@iupui.edu
Hochreiter
Amelia E
AE
Wright
Woodring E
WE
Shay
Jerry W
JW
D017931:DNA Primers; D005453:Fluorescence; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019098:Telomerase / Q000032:analysis*; D016615:Telomere / Q000235:genetics*; D020079:Terminal Repeat Sequences / Q000235:genetics
D017931:DNA Primers; D019098:Telomerase