title: A graphene-based platform for single nucleotide polymorphism (SNP) genotyping.
abstract: A facile, rapid, stable and sensitive approach for fluorescent detection of single nucleotide polymorphism (SNP) is designed based on DNA ligase reaction and π-stacking between the graphene and the nucleotide bases. In the presence of perfectly matched DNA, DNA ligase can catalyze the linkage of fluorescein amidite-labeled single-stranded DNA (ssDNA) and a phosphorylated ssDNA, and thus the formation of a stable duplex in high yield. However, the catalytic reaction cannot effectively carry out with one-base mismatched DNA target. In this case, we add graphene to the system in order to produce different quenching signals due to its different adsorption affinity for ssDNA and double-stranded DNA. Taking advantage of the unique surface property of graphene and the high discriminability of DNA ligase, the proposed protocol exhibits good performance in SNP genotyping. The results indicate that it is possible to accurately determine SNP with frequency as low as 2.6% within 40 min. Furthermore, the presented flexible strategy facilitates the development of other biosensing applications in the future.
authors.affiliation: Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education, China), School of Environmental Science and Technology, Dalian University of Technology, Dalian 116024, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Liu
authors.forename: Huimin
authors.initials: H
authors.lastname: Zhao
authors.forename: Shuo
authors.initials: S
authors.lastname: Chen
authors.forename: Hongtao
authors.initials: H
authors.lastname: Yu
authors.forename: Yaobin
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xie
authors.initials: X
authors.lastname: Quan
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods / Q000706:statistics & numerical data; D004247:DNA / Q000235:genetics; D011088:DNA Ligases; D004277:DNA, Single-Stranded / Q000235:genetics; D005453:Fluorescence; D005838:Genotype; D006108:Graphite; D020641:Polymorphism, Single Nucleotide; D013050:Spectrometry, Fluorescence
chemical_list: D004277:DNA, Single-Stranded; D006108:Graphite; D004247:DNA; D011088:DNA Ligases