Separation of mismatched DNA by using the affinity column immobilizing mismatch-binding ligands.
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.
Goto
Yuki
Y
Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, Kyoto 615-8510, Japan.
Kobori
Akio
A
Suda
Hitoshi
H
Nakatani
Kazuhiko
K
D020137:Base Pair Mismatch; D001483:Base Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D008024:Ligands; D009287:Naphthyridines / Q000737:chemistry; D009690:Nucleic Acid Conformation; D013997:Time Factors
D008024:Ligands; D009287:Naphthyridines; D004247:DNA