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4.5. Telomerase repeat amplification protocol (TRAP)
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The reaction mixture for assaying inhibition of human
telomerase (50 ll) contains 50 lM dNTPs, 0,4 lM
TSG4 primer and 1 ll of cell extract (prepared from
107 cultured human Neuroblastoma cells) in TRAP buff-
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of the cell extract. After 30 min of incubation at 30 ꢁC,
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evaluate PCR amplification efficiency.28
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Acknowledgments
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This research has been financially supported by
MIUR-FIRB 2001, MIUR-COFIN 2005 and by
Istituto Pasteur-Fondazione Cenci Bolognetti. Thanks
are due to S. Nasi and D. Annibali for having
provided the human Neuroblastoma cells for telome-
rase extract, to S. Bacchetti for the protocol for
telomerase extraction, to Dr. Luigi Rossetti for helpful
discussion and to Dr. Christoph Schultes for help in
the final revision of the manuscript. A financial contri-
bution by ARPA-Lazio funding for a research fellow-
ship to C. Mauriello is gratefully acknowledged.
23. Sonogashira, K.; Tohda, Y.; Hagihara, N. Tetrahedron
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Supplementary data
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Supplementary data associated with this article can
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