DdTRAP: A method for sensitive and precise quantification of telomerase activity

Andrew T. Ludlow; Dawne Shelton; Woodring E. Wright; Jerry W. Shay

doi:10.1007/978-1-4939-7778-9_29

DdTRAP: A method for sensitive and precise quantification of telomerase activity

Andrew T. Ludlow, Dawne Shelton, Woodring E. Wright, Jerry W. Shay

Research output: Chapter in Book/Report/Conference proceeding › Chapter

15 Scopus citations

Abstract

Telomerase is a cellular RNA template-dependent reverse transcriptase that adds telomere repeats to the 3′ ends of chromosomes. Telomerase is expressed almost universally in tumor cells (>85%) to maintain telomere length, thus providing the ability of tumor cells to avoid senescence and to have unlimited replication ability, one of the key hallmarks of cancer. ddTRAP (droplet digital Telomere Repeat Amplification Protocol) is a two-step assay with whole cell lysates that utilizes a telomerase-mediated primer extension followed by droplet digital PCR (ddPCR) detection of extended products. The adoptation of the TRAP assay to ddPCR has resulted in improved throughput, increased sensitivity and better repeatability of the TRAP assay. The protocol described below details our procedures for ddTRAP.

Original language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	513-529
Number of pages	17
DOIs	https://doi.org/10.1007/978-1-4939-7778-9_29
State	Published - 2018

Publication series

Name	Methods in Molecular Biology
Volume	1768
ISSN (Print)	1064-3745

Keywords

Droplet digital PCR
Telomerase activity

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

10.1007/978-1-4939-7778-9_29

Cite this

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title = "DdTRAP: A method for sensitive and precise quantification of telomerase activity",

abstract = "Telomerase is a cellular RNA template-dependent reverse transcriptase that adds telomere repeats to the 3′ ends of chromosomes. Telomerase is expressed almost universally in tumor cells (>85%) to maintain telomere length, thus providing the ability of tumor cells to avoid senescence and to have unlimited replication ability, one of the key hallmarks of cancer. ddTRAP (droplet digital Telomere Repeat Amplification Protocol) is a two-step assay with whole cell lysates that utilizes a telomerase-mediated primer extension followed by droplet digital PCR (ddPCR) detection of extended products. The adoptation of the TRAP assay to ddPCR has resulted in improved throughput, increased sensitivity and better repeatability of the TRAP assay. The protocol described below details our procedures for ddTRAP.",

keywords = "Droplet digital PCR, Telomerase activity",

author = "Ludlow, {Andrew T.} and Dawne Shelton and Wright, {Woodring E.} and Shay, {Jerry W.}",

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T2 - A method for sensitive and precise quantification of telomerase activity

AU - Ludlow, Andrew T.

AU - Shelton, Dawne

AU - Wright, Woodring E.

AU - Shay, Jerry W.

N1 - Publisher Copyright: © Springer Science+Business Media, LLC, part of Springer Nature 2018.

PY - 2018

Y1 - 2018

N2 - Telomerase is a cellular RNA template-dependent reverse transcriptase that adds telomere repeats to the 3′ ends of chromosomes. Telomerase is expressed almost universally in tumor cells (>85%) to maintain telomere length, thus providing the ability of tumor cells to avoid senescence and to have unlimited replication ability, one of the key hallmarks of cancer. ddTRAP (droplet digital Telomere Repeat Amplification Protocol) is a two-step assay with whole cell lysates that utilizes a telomerase-mediated primer extension followed by droplet digital PCR (ddPCR) detection of extended products. The adoptation of the TRAP assay to ddPCR has resulted in improved throughput, increased sensitivity and better repeatability of the TRAP assay. The protocol described below details our procedures for ddTRAP.

AB - Telomerase is a cellular RNA template-dependent reverse transcriptase that adds telomere repeats to the 3′ ends of chromosomes. Telomerase is expressed almost universally in tumor cells (>85%) to maintain telomere length, thus providing the ability of tumor cells to avoid senescence and to have unlimited replication ability, one of the key hallmarks of cancer. ddTRAP (droplet digital Telomere Repeat Amplification Protocol) is a two-step assay with whole cell lysates that utilizes a telomerase-mediated primer extension followed by droplet digital PCR (ddPCR) detection of extended products. The adoptation of the TRAP assay to ddPCR has resulted in improved throughput, increased sensitivity and better repeatability of the TRAP assay. The protocol described below details our procedures for ddTRAP.

KW - Droplet digital PCR

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DdTRAP: A method for sensitive and precise quantification of telomerase activity

Abstract

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