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| Zero-Background Helicase-Dependent Amplification and Its Application to Reliable Assay of Telomerase Activity in Cancer Cell by Eliminating Primer-Dimer Artifacts | |
| Chen, F; Zhang, DX; Zhang, Q; Zuo, XL; Fan, CH; Zhao, YX; Zhao, YX (reprint author), Xi An Jiao Tong Univ, Educ Minist, Sch Life Sci & Technol, Key Lab Biomed Informat Engn, Xianning West Rd, Xian 710049, Shaanxi, Peoples R China. | |
| 2016 | |
| Source Publication | CHEMBIOCHEM
 |
| ISSN | 1439-4227 |
| Volume | 17Issue:12Pages:1171-1176 |
| Subtype | 期刊论文 |
| Abstract | Primer-dimer artifacts resulting from unintended template-independent primer-primer interactions often hinder the specific amplification of nucleic acids. We demonstrate, for the first time, zero-background helicase-dependent amplification (HDA), with low concentrations of both ATP and dNTPs. This strategy achieved the reliable evaluation of telomerase activity in cancer cells by eliminating primer-dimer artifacts, which have plagued many previous methods with reduced specificity. We found that the performance of the telomerase assay by zero-background HDA was negatively affected by highly concentrated cellular proteins. This inhibitory effect is attributed to the binding of DNA templates to proteins, thus making them unavailable for polymerases. However, gold nanoparticles were demonstrated to highly attenuate such inhibition by abundant proteins, and to enhance the assay sensitivity and reliability when the reaction was performed with concentrated cell extracts. |
| Keyword | Helicase-dependent Amplification Nanoparticles Polymerase Chain Reaction Primer-dimer Artifacts Telomerase Activity Zero-background |
| DOI | 10.1002/cbic.201500605 |
| Indexed By | SCI |
| Language | 英语 |
| WOS ID | WOS:000379156400021 |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.sinap.ac.cn/handle/331007/26680 |
| Collection | 中科院上海应用物理研究所2011-2020年 |
| Corresponding Author | Zhao, YX (reprint author), Xi An Jiao Tong Univ, Educ Minist, Sch Life Sci & Technol, Key Lab Biomed Informat Engn, Xianning West Rd, Xian 710049, Shaanxi, Peoples R China. |
| Recommended Citation GB/T 7714 | Chen, F,Zhang, DX,Zhang, Q,et al. Zero-Background Helicase-Dependent Amplification and Its Application to Reliable Assay of Telomerase Activity in Cancer Cell by Eliminating Primer-Dimer Artifacts[J]. CHEMBIOCHEM,2016,17(12):1171-1176. |
| APA | Chen, F.,Zhang, DX.,Zhang, Q.,Zuo, XL.,Fan, CH.,...&Zhao, YX .(2016).Zero-Background Helicase-Dependent Amplification and Its Application to Reliable Assay of Telomerase Activity in Cancer Cell by Eliminating Primer-Dimer Artifacts.CHEMBIOCHEM,17(12),1171-1176. |
| MLA | Chen, F,et al."Zero-Background Helicase-Dependent Amplification and Its Application to Reliable Assay of Telomerase Activity in Cancer Cell by Eliminating Primer-Dimer Artifacts".CHEMBIOCHEM 17.12(2016):1171-1176. |
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| File Name/Size | DocType | Version | Access | License | ||
| Zero-Background Heli（1246KB） | 期刊论文 | 作者接受稿 | 开放获取 | CC BY-NC-SA | View Download | |
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