Abstract
Establishing low‐cost, high‐throughput, simple, and accurate single nucleotide polymorphism (SNP) genotyping techniques is beneficial for understanding the intrinsic relationship between individual genetic variations and their biological functions on a genomic scale. Here, a straightforward and reliable single‐molecule approach is demonstrated for precise SNP authentication by directly measuring the fluctuations in electrical signals in an electronic circuit, which is fabricated from a high‐gain field‐effect silicon nanowire decorated with a single hairpin DNA, in the presence of different target DNAs. By simply comparing the proportion difference of a probe‐target duplex structure throughout the process, this study implements allele‐specific and accurate SNP detection. These results are supported by the statistical analyses of different dynamic parameters such as the mean lifetime and the unwinding probability of the duplex conformation. In comparison with conventional polymerase chain reaction and optical methods, this convenient and label‐free method is complementary to existing optical methods and also shows several advantages, such as simple operation and no requirement for fluorescent labeling, thus promising a futuristic route toward the next‐generation genotyping technique.
Highlights
Introduction typing by usingDNA polymerase or ligase,[7] and enzyme mismatch cleavages.[8]
We have demonstrated a label-free and convenient approach for SNP discrimination through directly measuring the dynamic electrical fluctuation of probe-target duplex hybridization in a single hairpin DNA-decorated silicon nanowire (SiNW) field-effect transistor (FET) electronic circuit
We accomplished the allele-specific detection by comparing the proportion differences of the probe-target duplex structure during the entire process, which can be strengthened by the dynamic differences including the mean lifetime and transition probability of the duplex conformation
Summary
SiNW-based single-molecule electrical biosensors (Figure 1a) were prepared by using a well-developed strategy.[15e,16] The detailed process of device fabrication and DNA attachment is provided in the Experimental Section. 78.5 ± 1.8 20.2 ± 1.8 1.3 ± 0.6 3.6 ± 1.2 90.0 ± 0.8 6.4 ± 0.6 8.3 ± 1.8 72.0 ± 2.6 19.7 ± 1.8 1.6 ± 0.1 90.5 ± 1.0 7.9 ± 1.0 of the lack of essential NHS-esterification, demonstrated that when the blank device was exposed to PBS buffer, 1 × 10−6 m WT-C, MT-A, MT-G, MT-T, and Non-C DNAs solutions, respectively, the current recordings ΔID(t) exhibited no particular fluctuations with only a Gaussian distribution dominated by 1/f noise (Figure S4, Supporting Information) These results eliminate the possibilities of nonspecific surface absorption of either hairpin DNAs or ions in the electrolytic solution, support that the three-level current oscillations originated from the intrinsic behaviors of hairpin DNA hybridization with the targets.
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