Nonradioactive detection of telomerase activity using the telomeric repeat amplification protocol |
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| Authors: | Herbert Brittney-Shea Hochreiter Amelia E Wright Woodring E Shay Jerry W |
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| Institution: | Department of Medical and Molecular Genetics, Indiana University Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana 46202-5251, USA. brherber@iupui.edu |
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| Abstract: | 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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