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Nonradioactive detection of telomerase activity using the telomeric repeat amplification protocol

Nature Protocols volume 1, pages 1583–1590 (2006)Cite this article

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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Figure 1: Analysis of telomerase activity using Cy5-TRAP.
Figure 2: Real-time Q-TRAP analysis of telomerase activity: preparation of the standard curve.

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Acknowledgements

We thank P. Erickson and E. Badenhop for technical assistance. This work was supported by the Indiana University Cancer Center and the Indiana Genomics Initiative (INGEN). INGEN of Indiana University is supported in part by Lilly Endowment Inc.

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Authors and Affiliations

  1. Department of Medical and Molecular Genetics, Indiana University Cancer Center, Indiana University School of Medicine, Indianapolis, 46202-5251, Indiana, USA

    Brittney-Shea Herbert & Amelia E Hochreiter

  2. Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, 75390-9039, Texas, USA

    Woodring E Wright & Jerry W Shay

Authors
  1. Brittney-Shea Herbert
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  2. Amelia E Hochreiter
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  3. Woodring E Wright
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  4. Jerry W Shay
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Correspondence to Brittney-Shea Herbert.

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JWS and WEW consult for, hold stock in, and are members of the scientific advisory board of Geron Corporation. Other authors declare that they have no competing financial interests.

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Herbert, BS., Hochreiter, A., Wright, W. et al. Nonradioactive detection of telomerase activity using the telomeric repeat amplification protocol. Nat Protoc 1, 1583–1590 (2006). https://doi.org/10.1038/nprot.2006.239

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