This Protocol is listed in the following Categories:
Cell and developmental biology

Author(s): Brittney-Shea Herbert, Amelia E Hochreiter, Woodring E Wright & Jerry W Shay
Affiliation(s): Department of Medical and Molecular Genetics, Indiana University Cancer Center, Indiana University School of Medicine
DOI: 10.1038/nprot.2006.239

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).

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