Graphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching

Abstract Aiming at improved specificity, nanoparticle assisted polymerase chain reaction (PCR) has been widely studied and shown to improve PCR. However, the reliability and mechanism of this method are still controversial. Here, we demonstrated that 1 μg/mL of graphene oxide (GO) effectively enhanc...

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Autores principales:	Yuanyuan Wang, Fengbang Wang, Hailin Wang, Maoyong Song
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e19f70c33a0841d5a38bba6b608dd69a
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spelling	oai:doaj.org-article:e19f70c33a0841d5a38bba6b608dd69a2021-12-02T15:04:55ZGraphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching10.1038/s41598-017-16836-x2045-2322https://doaj.org/article/e19f70c33a0841d5a38bba6b608dd69a2017-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-16836-xhttps://doaj.org/toc/2045-2322Abstract Aiming at improved specificity, nanoparticle assisted polymerase chain reaction (PCR) has been widely studied and shown to improve PCR. However, the reliability and mechanism of this method are still controversial. Here, we demonstrated that 1 μg/mL of graphene oxide (GO) effectively enhances the specificity of the error-prone multi-round PCR. Mismatched primers were designed as interference to produce nonspecific products when the same amounts of matched and mismatched primers were added into semi-multiplex PCR. It was found that GO can enhance specificity by suppressing the amplification of mismatched primers. We monitored the primer-template-polymerase-GO interactions involved in the PCR using a capillary electrophoresis/laser-induced fluorescence polarization (CE-LIFP) assay. The results showed that the addition of GO promoted the formation of a matched primer-template complex, but suppressed the formation of a mismatched primer-template complex during PCR, suggesting that interactions between the primers and GO play an essential role. Furthermore, we successfully amplified the FOXL2 gene from PEGFP-N1 vectors using GO to eliminate the nonspecific products in PCR. Taken together, these results suggest that the GO can be used as an efficient additive for improving the conventional PCR system.Yuanyuan WangFengbang WangHailin WangMaoyong SongNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yuanyuan Wang Fengbang Wang Hailin Wang Maoyong Song Graphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching
description	Abstract Aiming at improved specificity, nanoparticle assisted polymerase chain reaction (PCR) has been widely studied and shown to improve PCR. However, the reliability and mechanism of this method are still controversial. Here, we demonstrated that 1 μg/mL of graphene oxide (GO) effectively enhances the specificity of the error-prone multi-round PCR. Mismatched primers were designed as interference to produce nonspecific products when the same amounts of matched and mismatched primers were added into semi-multiplex PCR. It was found that GO can enhance specificity by suppressing the amplification of mismatched primers. We monitored the primer-template-polymerase-GO interactions involved in the PCR using a capillary electrophoresis/laser-induced fluorescence polarization (CE-LIFP) assay. The results showed that the addition of GO promoted the formation of a matched primer-template complex, but suppressed the formation of a mismatched primer-template complex during PCR, suggesting that interactions between the primers and GO play an essential role. Furthermore, we successfully amplified the FOXL2 gene from PEGFP-N1 vectors using GO to eliminate the nonspecific products in PCR. Taken together, these results suggest that the GO can be used as an efficient additive for improving the conventional PCR system.
format	article
author	Yuanyuan Wang Fengbang Wang Hailin Wang Maoyong Song
author_facet	Yuanyuan Wang Fengbang Wang Hailin Wang Maoyong Song
author_sort	Yuanyuan Wang
title	Graphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching
title_short	Graphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching
title_full	Graphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching
title_fullStr	Graphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching
title_full_unstemmed	Graphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching
title_sort	graphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/e19f70c33a0841d5a38bba6b608dd69a
work_keys_str_mv	AT yuanyuanwang grapheneoxideenhancesthespecificityofthepolymerasechainreactionbymodifyingprimertemplatematching AT fengbangwang grapheneoxideenhancesthespecificityofthepolymerasechainreactionbymodifyingprimertemplatematching AT hailinwang grapheneoxideenhancesthespecificityofthepolymerasechainreactionbymodifyingprimertemplatematching AT maoyongsong grapheneoxideenhancesthespecificityofthepolymerasechainreactionbymodifyingprimertemplatematching
_version_	1718388988338765824

Graphene oxide enhances the specificity of the polymerase chain reaction by modifying primer-template matching

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