کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1212143 | 1494055 | 2015 | 6 صفحه PDF | دانلود رایگان |

• We developed a method for telomerase activity by microchip electrophoresis.
• The mixed gel of hydroxypropyl methylcellulose and polyethylene oxide was used.
• Detection of PCR products was possible in as few as 20 cycles.
• A sample as small as a single cell could detected in the short time of 100 s.
• Two human cancer cell lines were analyzed satisfactorily using this method.
Telomerase participates in malignant transformation or immortalization of cells and thus has attracted attention as an anticancer drug target and diagnostic tumor marker. The telomeric repeat amplification protocol (TRAP) and improved TRAP methods (TRAP-fluorescence, TRAP-hybridization, etc.) are widely used forms of this telomerase assay. However, these approaches generally employ acrylamide gel electrophoresis after amplification of telomeric repeats by polymerase chain reaction (PCR), making these TRAP methods time consuming and technically demanding. In this study we developed a novel telomerase assay using microchip electrophoresis for rapid and highly sensitive detection of telomerase activity in cancer cells. The mixed gel of 0.8% hydroxypropyl methylcellulose (HPMC) and 0.3% polyethylene oxide (PEO) with SYBR Gold (fluorescent reagent) was used for microchip electrophoresis. As a result, the product amplified by a telomerase-positive cell could be measured in one cell per assay and detected with high reproducibility (CV = 0.67%) in the short time of 100 s.
Journal: Journal of Chromatography B - Volumes 993–994, 1 July 2015, Pages 14–19