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영어

We present a simple and efficient method for specificity improvement of SNPs detection that combines PNA microarray with single-stranded DNA specific nuclease (S1 nuclease). When DNA is perfectly matched with PNA, the DNA/PNA duplexes are not digested by S1 nuclease as the PNA protects the DNA from the nuclease. However, DNA/PNA duplexes including one-base
mismatch is efficiently hydrolyzed by the nuclease. This phenomenon was applied to the PNA microarray for single nucleotide polymorphisms (SNPs) detection. With Cy3-labeled target oligonucleotide hybridization to PNA microarray, non specific binding to mismatched PNA probe shows unexpected fluorescent intensity causing low specificity. Though, after treatment of S1
nuclease on PNA microarray, the intensity ratio between perfect matched and one-base mismatched case sharply increased. Therefore, we expect that combining PNA microarray with S1 nuclease will be a powerful tool in the development of accurate and high throughput SNPs detection.