title: The addition of a spin column step in the telomeric repeat application protocol removes telomerase inhibitors.
abstract: Telomerase activity in cancer cells is commonly analyzed by a polymerase chain reaction (PCR)-based assay termed the telomeric repeat amplification protocol (TRAP). However, nonspecific inhibition of Taq polymerase during the PCR step is frequently observed in inhibitor analysis or drug screening. Thus, the removal of excess inhibitors prior to PCR is an essential step for the proper evaluation of telomerase inhibitory effects. Here, a size exclusion spin column was applied to remove small molecular weight inhibitors from the telomerase extension products. The spin column-added protocol, termed sTRAP, provides a more reliable estimation of the inhibitory effects of telomerase activity.
authors: Institute of Biopharmaceutical Sciences, National Yang-Ming University, Taipei, Taiwan.
authors: Ying-Chieh
authors: YC
authors: Chen
authors: Institute of Biochemistry and Molecular Biology, National Taiwan University College of Medicine, Taipei, Taiwan.
authors: Fong-Chun
authors: FC
authors: Huang
authors: Institute of Biopharmaceutical Sciences, National Yang-Ming University, Taipei, Taiwan; Institute of Biochemistry and Molecular Biology, National Taiwan University College of Medicine, Taipei, Taiwan. Electronic address: jingjerlin@ntu.edu.tw.
authors: Jing-Jer
authors: JJ
authors: Lin
mesh_terms: D045744:Cell Line, Tumor; D003911:Dextrans / Q000737:chemistry; D004354:Drug Screening Assays, Antitumor; D004791:Enzyme Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D054852:Small Molecule Libraries / Q000302:isolation & purification / Q000494:pharmacology; D019914:Taq Polymerase / Q000037:antagonists & inhibitors / Q000378:metabolism; D019098:Telomerase / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D003911:Dextrans; D004791:Enzyme Inhibitors; D054852:Small Molecule Libraries; C025614:sephadex; D019914:Taq Polymerase; D019098:Telomerase
keywords: Anti-cancer; Inhibitor; PCR; TRAP; Telomerase