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Info. Vol.9 - No.2 (2015.06.20)
Title Multispot Array Combined with S1 Nuclease-Mediated Elimination of Unpaired Nucleotides
Authors Seung Min Yoo1,??/sup>, Dong Min Kim2,??/sup> & Sang Yup Lee1,3,*
Institutions 1Department of Chemical and Biomolecular Engineering (BK21 plus program), BioProcess Engineering Research Center, and Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, KAIST, Daejeon 305-701, Korea
2Center for Applied Life Science, Hanbat National University, Daejeon 305-719, Korea
3The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Alle 6, H첩rsholm, Denmark
Abstract The accurate detection of mismatched base pairs is critical to many DNA hybridization-based applications in basic research and diagnostics. We herein demonstrate that mismatched DNAs on a multispot array can be accurately detected in a multiplexed way by employing the S1 nuclease-based mismatched base pair-specific cleavage system. After the optimization of the reaction condition, mismatched DNAs present in various pathogenic bacteria and genetic disorders could be successfully detected with stable hybridization signals regardless of the position of the fluorescent label relative to the probe-target duplex. This technique of performing S1 nuclease-mediated cleavage on a multispot array offers high specificity and high-throughput detection of mismatched DNAs. It is expected that this assay system will prove useful for single-assay genotyping and/or the diagnosis of various diseases and pathogens.
Keyword S1 nuclease, Enzymatic cleavage, Multispot array, DNA, Single mutation
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