التفاصيل البيبلوغرافية
| العنوان: |
Combined SYBR Green Real-Time with Telomeric Repeat Amplification Protocol (RQ-TRAP) to Detect Telomerase Activity. (English) |
| المؤلفون: |
MA wenqing, LIAN Fuzhi, WANG Jinquan, YANG Lei |
| المصدر: |
Tianjin Medical Journal; Aug2014, Vol. 42 Issue 8, p746-748, 3p |
| مصطلحات موضوعية: |
TELOMERASE genetics, GENE amplification, POLYMERASE chain reaction, ENZYME-linked immunosorbent assay, PHYSIOLOGICAL effects of proteins |
| مستخلص: |
Objective To establish methodology to detect telomerase activity based on real-time quantitative PCR technique combined with telomeric repeat amplification protocol (TRAP). Methods RQ-TRAP system was developed by combining real-time quantitative PCR technique with conventional TRAP method. Telomerase activity was assessed and compared by RQ-TRAP assay and TRAP connected with enzyme-linked immunosorbent assay (TRAP-ELISA) respectively in 12 kinds of cells. Results The RQ-TRAP method was both accurate and specified in measuring telomerase activity in a series dilution of protein extracts from 293T cells. The sensitivity of this method was 8 cells and the amplification efficiency was 98.92%. Telomerase activity was not detected in negative control group. Statistical analysis revealed a strong correlation between the two assays (r²=0.762 5). Conclusion The feasibility of RQ-TRAP was proved in this article. Compared with TRAP-ELISA, RQ-TRAP has many advantages. Apart from sample extraction and real-time PCR cycling, no other extra time-consuming steps are needed for telomerase quantification; RQ-TRAP is less costly and more rapid and reliable than TRAP-ELISA for quantification of telomerase activity and it also support high throughput. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
