Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009:523:383-94.
doi: 10.1007/978-1-59745-190-1_25.

Non-radioactive assay methods for the assessment of telomerase activity and telomere length

Partha P Banerjee  1 , Shankar Jagadeesh
Affiliations

Non-radioactive assay methods for the assessment of telomerase activity and telomere length

Partha P Banerjee et al. Methods Mol Biol. 2009.

Abstract

The telomeric repeat amplification protocol (TRAP) assay is a highly sensitive PCR based assay and is an important tool for understanding the role of telomerase in cancer. This assay measures an enzymatic activity where the amount of target is dependent upon the activity of the enzyme. This protocol consists of two steps: first, telomeric repeats are added to the substrate by the enzyme and second, the extended products will be amplified by Taq-DNA polymerase. The amplified TRAP assay products will be separated on 10% native PAGE and detected by SYBR Green I dye.

PubMed Disclaimer

Figures

Fig. 25.1
Fig. 25.1
A flow chart depicting the various steps in the TRAP assay.
Fig. 25.2
Fig. 25.2
Determination of telomerase activity in prostate cancer cells (LNCaP) with or without treatments of a telomerase inhibitor for 3 days in culture. Telomerase was extracted from cell pellets and subjected to TRAP assay. NC, negative control using lysis buffer only.
Fig. 25.3
Fig. 25.3
Determination of telomere length in prostate cancer cells (LNCaP) with or without the treatment of a telomerase inhibitor for 3 days. N, normal (untreated) LNCaP cells; mean telomere length is 2.5 kb; TI, LNCaP cells treated with telomerase inhibitor; mean telomere length is 1.5 kb. MWM, molecular weight marker.
See this image and copyright information in PMC

References

    1. Greider CW, Blackburn EH. A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis. Nature. 1989;337:331–337. - PubMed
    1. Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL, Cov-iello GM, Wright WE, Weinrich SL, Shay JW. Specific association of human telomerase activity with immortal cells and cancer. Science. 1994;266:2011–2015. - PubMed
    1. Harley CB, Villeponteau B. Telomeres and telomerase in aging and cancer. Curr. Opin. Genet. Dev. 1995;5(5):249–255. - PubMed
    1. Kilian A, Bowtell DD, Abud HE, Hime GR, Venter DJ, Keese PK, Duncan EL, Reddel RR, Jefferson RA. Isolation of a candidate human telomerase catalytic subunit gene, which reveals complex splicing patterns in different cell types. Hum. Mol. Genet. 1997;6:2011–2019. - PubMed
    1. Feng J, Funk WD, Wang SS, Weinrich SL, Avilion AA, Chiu CP, Adams RR, Chang E, Allsopp RC, Yu J, et al. The RNA component of human telomerase. Science. 1995;269:1236–1241. - PubMed

LinkOut - more resources