title: A fluorescence method for detection of DNA and DNA methylation based on graphene oxide and restriction endonuclease HpaII.
abstract: DNA methylation plays an important role in many biological events and is associated with various diseases. Most traditional methods for detection of DNA methylation are based on the complex and expensive bisulfite method. In this paper, we report a novel fluorescence method to detect DNA and DNA methylation based on graphene oxide (GO) and restriction endonuclease HpaII. The skillfully designed probe DNA labeled with 5-carboxyfluorescein (FAM) and optimized GO concentration keep the probe/target DNA still adsorbed on the GO. After the cleavage action of HpaII the labeled FAM is released from the GO surface and its fluorescence recovers, which could be used to detect DNA in the linear range of 50 pM-50 nM with a detection limit of 43 pM. DNA methylation induced by transmethylase (Mtase) or other chemical reagents prevents HpaII from recognizing and cleaving the specific site; as a result, fluorescence cannot recover. The fluorescence recovery efficiency is closely related to the DNA methylation level, which can be used to detect DNA methylation by comparing it with the fluorescence in the presence of intact target DNA. The method for detection of DNA and DNA methylation is simple, reliable and accurate.
authors.affiliation: Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Chemistry and Chemical Engineering, Southeast University, Jiangning District, Nanjing 211189, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.affiliation: Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Chemistry and Chemical Engineering, Southeast University, Jiangning District, Nanjing 211189, PR China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Gao
authors.affiliation: Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Chemistry and Chemical Engineering, Southeast University, Jiangning District, Nanjing 211189, PR China.
authors.forename: Yanxiang
authors.initials: Y
authors.lastname: Xiong
authors.affiliation: Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Chemistry and Chemical Engineering, Southeast University, Jiangning District, Nanjing 211189, PR China.
authors.forename: Yuanjian
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Chemistry and Chemical Engineering, Southeast University, Jiangning District, Nanjing 211189, PR China. Electronic address: liusq@seu.edu.cn.
authors.forename: Songqin
authors.initials: S
authors.lastname: Liu
authors.affiliation: Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Jiangning District, Nanjing 211189, PR China. Electronic address: yppu@seu.edu.cn.
authors.forename: Yuepu
authors.initials: Y
authors.lastname: Pu
mesh_terms: D004247:DNA / Q000032:analysis*; D019175:DNA Methylation; D015342:DNA Probes / Q000737:chemistry; D019003:Deoxyribonuclease HpaII / Q000378:metabolism*; D005452:Fluoresceins / Q000737:chemistry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D006108:Graphite / Q000737:chemistry*; D057230:Limit of Detection; D013050:Spectrometry, Fluorescence
chemical_list: D015342:DNA Probes; D005452:Fluoresceins; D005456:Fluorescent Dyes; C045132:4-carboxyfluorescein; D006108:Graphite; D004247:DNA; D019003:Deoxyribonuclease HpaII
keywords: DNA methylation; Fluorescence; Graphene oxide; HpaII; Restriction endonuclease