Telomerase assay using biotinylated-primer extension and magnetic separation of the products

Daekyu Sun, Laurence Hurley, Daniel D. Von Hoff

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Human telomerase, a ribonucleoprotein enzyme, is known to be associated with immortalized cancer cells but is absent in most normal tissues. Thus, telomerase appears to be an attractive new target for anticancer agents and an important diagnostic marker of human cancers. Here, we describe an improved telomerase assay method based on the Dynabead® biomagnetic separation theory. In this method, 5'-biotinylated (TTAGGG)3 was used as a primer for the telomerase reaction. Telomerase reaction products were then immobilized on streptavidin-coated Dynabeads and washed intensively to eliminate excess [α32P]dGTP. Using this method, without the amplification of telomerase reaction products by the PCR, we were able to quantitatively detect telomerase activity in human HeLa cell extracts equivalent to between 200-500 cells. This method is anticipated to be useful for the measurement of telomerase activity in various tumor cells, for assessing potential telomerase and for understanding the biochemical aspects of the telomerase reaction.

Original languageEnglish (US)
Pages (from-to)1046-1051
Number of pages6
JournalBioTechniques
Volume25
Issue number6
StatePublished - 1998
Externally publishedYes

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Magnetic separation
Telomerase
Assays
Reaction products
Cells
Neoplasms
Ribonucleoproteins
Streptavidin
Cell Extracts
HeLa Cells
Human Activities
Antineoplastic Agents
Amplification
Tumors
Tissue
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Telomerase assay using biotinylated-primer extension and magnetic separation of the products. / Sun, Daekyu; Hurley, Laurence; Von Hoff, Daniel D.

In: BioTechniques, Vol. 25, No. 6, 1998, p. 1046-1051.

Research output: Contribution to journalArticle

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