Increased sensitivity and reproducibility of TRAP assay by avoiding direct primers interaction.
Telomeric Repeat Amplification Protocol (TRAP) is a sensitive procedure to measure telomerase activity in small samples of cell or tissue extracts. Due to the strict correlation between high levels of telomerase activity and neoplastic transformation, TRAP assay could provide an important diagnostic marker of malignancy. Although the original TRAP assay is very sensitive and some improvements have been described, occasional artifacts still persist in the modified procedures. Here we describe how changes in the sequence of the primer used for the amplification step enhance the reproducibility and sensitivity in the TRAP assay.
Falchetti
M L
ML
Dipartimento di Medicina Sperimentale, University of l'Aquila, Rome, Italy.
Levi
A
A
Molinari
P
P
Verna
R
R
D'Ambrosio
E
E
D000818:Animals; D002457:Cell Extracts; D002460:Cell Line; D017931:DNA Primers; D006801:Humans; D008579:Meningioma / Q000201:enzymology; D051379:Mice; D009000:Monocytes; D016716:PC12 Cells; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019098:Telomerase / Q000378:metabolism*
D002457:Cell Extracts; D017931:DNA Primers; D019098:Telomerase