|Title:||The addition of a spin column step in the telomeric repeat application protocol removes telomerase inhibitors||Authors:||Chen Y.-C.
|Keywords:||Anti-cancer; Inhibitor; PCR; Telomerase; TRAP||Issue Date:||2015||Publisher:||Academic Press Inc.||Journal Volume:||478||Start page/Pages:||49-51||Source:||Analytical Biochemistry||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. ? 2015 Elsevier Inc. All rights reserved.
|ISSN:||0003-2697||DOI:||10.1016/j.ab.2015.03.020||SDG/Keyword:||Diseases; Amplification protocols; Anti-cancer; Application protocols; Inhibitor; Inhibitor analysis; Telomerase; Telomerase activity; TRAP; Polymerase chain reaction; Taq polymerase; telomerase; telomerase inhibitor; dextran; enzyme inhibitor; molecular library; sephadex; Taq polymerase; telomerase; antineoplastic activity; Article; assay; drug screening; enzyme activity; polymerase chain reaction; priority journal; telomeric repeat amplification protocol; antagonists and inhibitors; chemistry; human; isolation and purification; metabolism; molecular library; pharmacology; procedures; tumor cell line; Cell Line, Tumor; Dextrans; Drug Screening Assays, Antitumor; Enzyme Inhibitors; Humans; Polymerase Chain Reaction; Small Molecule Libraries; Taq Polymerase; Telomerase
|Appears in Collections:||生物化學暨分子生物學科研究所|
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