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Single nucleotide polymorphism detection method by temperature-gradient affinity chromatography using a single-stranded oligo-DNA coupled column.

Tomohisa Katsuda·Ken Nishijima·Mitsumasa Kamura·Yasushi Nishiwada·Shigeo Katoh
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Abstract

We developed an affinity chromatographic method for simple single nucleotide polymorphism (SNP) detection by use of a single-stranded DNA-coupled column and temperature gradient elution, utilizing the difference in thermal stability between hybridized double-stranded DNAs with and without mismatched base-pairs in the course of temperature gradient elution. We studied experimentally and theoretically the elution behavior of DNAs with and without SNPs in this chromatography and proposed a numerical calculation method based on a thermodynamic dissociation model. The effects of the column volume, flow rate of eluent and heating rate of the column on elution profiles were clarified. For designing DNA ligands, mismatched base-pair positions favorable for detection of SNPs were also explored by use of hybridized DNAs coding a part of the human TP53 gene.

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Base Pair MismatchBase SequenceChromatography, AffinityDNA, Single-StrandedNucleic Acid DenaturationPolymorphism, Single NucleotideTemperatureJournal Article

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Tomohisa Katsuda, Ken Nishijima, Mitsumasa Kamura, Yasushi Nishiwada, Shigeo Katoh. 2006-02-14. Single nucleotide polymorphism detection method by temperature-gradient affinity chromatography using a single-stranded oligo-DNA coupled column.. https://doi.org/10.1016/j.chroma.2006.01.062

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