期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 388, 期 5-6, 页码 1165-1173出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-007-1323-y
关键词
mismatch binding ligand; affinity chromatography; mutation detection; melting temperature; single nucleotide polymorphism
Mismatch binding molecules (MBLs), strongly and selectively bound to the mismatched base pair in duplex DNA, were immobilized on Sepharose. Three MBL-Sepharose columns were prepared with three MBLs, naphthyridine dimer (ND), naphthyridine-azaquinolone (NA), and aminonaphthyridine dimer (amND), which exhibited different binding profiles to the mismatched base pairs. These three MBL-Sepharose columns showed characteristic elution profiles for DNA duplexes containing mismatched base pairs. The ND-Sepharose column separated the G-G and G-A mismatched DNA from fully matched duplexes. The NA-Sepharose column separated the A-A and G-A mismatched DNA from other DNA duplexes. The amND-Sepharose column separated the C-C mismatched DNA. These chromatographic profiles were very consistent with the binding preference of each MBL. By changing the elution conditions from sodium hydroxide to sodium chloride, MBL-Sepharose columns were also able to separate the mismatched DNA that weakly bound to the MBL from fully matched DNA duplex.
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