Premature termination of telomeric extension-PCR for detection of telomerase activity.
In this article, we report a simple, rapid, and efficient method to detect telomerase activity: the premature termination of telomeric extension-PCR (PTEP). Similar to the telomeric repeat amplification protocol (TRAP), this method is based on PCR amplification following the in vitro telomerase reaction, while the in vitro telomerase reaction here is prematurely, rather than randomly, terminated. Apart from this, the telomeric extension products are used as initial primers, instead of as templates, to trigger the amplification with a specially constructed plasmid DNA as the template that cannot be directly amplified with the telomerase primer. The end product is a specific 159-bp DNA fragment that reflects telomerase activity. Because its product can be clearly identified with routine agarose gel electrophoresis and ethidium bromide staining, PTEP allows even lesser-equipped laboratories to easily detect telomerase activity.
Chen
Ran
R
State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P.R. China. rachen1970@yahoo.com
Qian
Ji
J
Wang
Lijuan
L
Mao
Yu-min
YM
D002460:Cell Line; D045744:Cell Line, Tumor; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004789:Enzyme Activation; D020869:Gene Expression Profiling / Q000379:methods*; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007938:Leukemia / Q000201:enzymology*; D008175:Lung Neoplasms / Q000201:enzymology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012016:Reference Values; D019098:Telomerase / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*
D019098:Telomerase