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Comparison of the telomeric repeat amplification protocol (TRAP) to the new TRAP-eze telomerase detection kit

  • Biotechnology
  • Published:
Methods in Cell Science

Abstract

The ribonucleoprotein, telomerase, is believed to be responsible for the maintenance of telomere length in immortal and cancer cells. A PCR-based assay for the detection of telomerase activity (TRAP assay: telomeric repeat amplification protocol) was developed, allowing fast and efficient detection of telomerase activity when sample amounts are limiting. Of the thousands of primary human tumors examined using the TRAP assay, almost 90% have been shown to exhibit telomerase activity. Thus, for the early detection of cancer and for the rapid screening of compounds and drugs in cancer therapeutics, methods for the detection of telomerase activity are rapidly emerging. The recently developed TRAP-ezeTM kit from Oncor, Inc. gives increased sensitivity with decrease sample processing time, allowing improved detection of telomerase activity in a large number of samples. In the present study, we have addressed some of the technical aspects and limitations of critical importance for reproducibility, reliability, and linearity of the standard TRAP assay and the TRAP-ezeTM kit using cell culture and clinical materials.

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Abbreviations

RP:

modified reverse primer (CX) for PCR amplification of telomerase products

AEBSF:

4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride

CHAPS:

3-[(3-cholamidopropyl)dimethylammoniol-1-propanesulfonate

CX:

reverse primer for PCR amplification of telomerase products for standard TRAP

DEPC:

diethyl pyrocabonate

EGTA:

ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid

HPLC:

high pressure liquid chromatography

ITAS:

internal telomerase assay standard

PCR:

polymerase chain reaction

T4g32 protein:

T4 gene 32 protein

TRAP:

telomeric repeat amplification protocol

TRF:

terminal restriction fragment

TS:

primer that serves as a substrate for telomerase

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Authors and Affiliations

  1. The Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, USA

    Shawn E. Holt, James C. Norton, Woodring E. Wright & Jerry W. Shay

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  1. Shawn E. Holt
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  2. James C. Norton
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  3. Woodring E. Wright
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  4. Jerry W. Shay
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Correspondence to Jerry W. Shay.

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Holt, S.E., Norton, J.C., Wright, W.E. et al. Comparison of the telomeric repeat amplification protocol (TRAP) to the new TRAP-eze telomerase detection kit. Methods Cell Sci 18, 237–248 (1996). https://doi.org/10.1007/BF00132889

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