Detection of telomerase activity in human cells and tumors by a telomeric repeat amplification protocol (TRAP)

Mieczyslaw A. Piatyszek, Nam W. Kim, Scott L. Weinrich, Keiko Hiyama, Eiso Hiyama, Woodring E. Wright, Jerry W. Shay

Research output: Contribution to journalArticle

387 Scopus citations

Abstract

The association of human telomerase activity with an indefinite replicative capacity of cells in vitro and advanced tumors in vivo is gaining wide support. The increasing interest in studying various aspects of telomerase expression in cancer required the development of a sensitive and reliable protocol for the extraction and detection of telomerase activity in cell culture material, and from small tissue samples obtained from biopsy, surgical reaction of tumors, and autopsy. Recently a novel procedure for the extraction and detection of telomerase activity was developed (Science 1994; 266: 2011-2015) which resulted in an estimated 104 fold improvement in detectability compared with previous methods. The described procedures not only dramatically increased sensitivity but also allowed fast and efficient detection of telomerase activity in a large number of samples. A number of technical aspects which are of eritical importance for reproducibility and reliability of this assay using clinical material are addressed in this report. In addition, new methods to perform telomerase assays without the use of radioisotopes are described.

Original languageEnglish (US)
Pages (from-to)1-15
Number of pages15
JournalMethods in Cell Science
Volume17
Issue number1
DOIs
StatePublished - Mar 1 1995

Keywords

  • Cancer
  • Cellular immortality
  • Telomerase
  • Telomeres

ASJC Scopus subject areas

  • Cell Biology

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