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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 PublicationCHEMBIOCHEM
ISSN1439-4227
Volume17Issue:12Pages:1171-1176
Subtype期刊论文
AbstractPrimer-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.
KeywordHelicase-dependent Amplification Nanoparticles Polymerase Chain Reaction Primer-dimer Artifacts Telomerase Activity Zero-background
DOI10.1002/cbic.201500605
Indexed BySCI
Language英语
WOS IDWOS:000379156400021
Citation statistics
Cited Times:7[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.sinap.ac.cn/handle/331007/26680
Collection中科院上海应用物理研究所2011-2018年
Corresponding AuthorZhao, 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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