Molecular basis of artifacts in the detection of telomerase activity and a modified primer for a more robust 'TRAP' assay | Zendy

Guido Krupp | Zendy; Klaus-Stefan Kühne | Zendy; Sandra Tamm | Zendy; Wolfram Klapper | Zendy; Klaus Heidorn | Zendy; A. Rott | Zendy; Reza Parwaresch | Zendy

Open Access

Molecular basis of artifacts in the detection of telomerase activity and a modified primer for a more robust 'TRAP' assay

Author(s) -

Guido Krupp,

Klaus-Stefan Kühne,

Sandra Tamm,

Wolfram Klapper,

Klaus Heidorn,

A. Rott,

Reza Parwaresch

Publication year - 1997

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/25.4.919

Subject(s) - telomerase , biology , primer (cosmetics) , telomere , trap (plumbing) , microbiology and biotechnology , polymerase chain reaction , reverse transcriptase , somatic cell , computational biology , genetics , dna , gene , chemistry , organic chemistry , environmental engineering , engineering

Human somatic cells have essentially no telomerase activity. Telomerase is linked to tumor genesis and is a valuable marker for malignant growth. Extreme paucity of the enzyme neccessitated development of a PCR-based assay, 'telomeric repeat amplification protocol' (TRAP). Unfortunately, this method is not without difficulties. Amplification products are not related to the size of the amplified telomerase products. Furthermore, false positive results can occur, and careful control of reaction conditions is crucial. We analyzed in detail the molecular basis of artifacts. Based on these data, reverse PCR primer was changed and both problems in the TRAP assay were eliminated.

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