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Separation of mismatched DNA by using the affinity column immobilizing mismatch-binding ligands
Yuki Goto1, Akio Kobori, Hitoshi Suda
1Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, Kyoto 615-8510, Japan.
Abstract:
Mismatch binding ligands (MBL), ND and NA, bind to DNA containing mismatch base pairs. These MBLs were immobilized to NHS-activated affinity column via amine linker. Affinity chromatographic analyses of mismatched DNAs by using the ND-immobilized column showed clear separation of G-G and G-A from other mismatched DNAs. The results of the NA-immobilized column also showed good separation of A-A, G-A, G-G, A-C from other mismatched DNAs. The order of mobility of mismatched duplexes on MBL-immobilized column was in good agreement with that of the binding affinity in a solution phase obtained by the melting temperature measurement.
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