Capillary Electrophoretic Discrimination of Single Nucleotide Polymorphisms Using an Oligodeoxyribonucleotide-polyacrylamide Conjugate as a Pseudo-immobilized Affinity Ligand: Optimum Ligand Length Predicted by the Melting Temperature Values

Accession number;05A0095062
Title;Capillary Electrophoretic Discrimination of Single Nucleotide Polymorphisms Using an Oligodeoxyribonucleotide-polyacrylamide Conjugate as a Pseudo-immobilized Affinity Ligand: Optimum Ligand Length Predicted by the Melting Temperature Values
Author; MISHIMA K (Riken (inst. Of Physical And Chemical Res.), Wako, Jpn) TAKARADA T (Riken (inst. Of Physical And Chemical Res.), Wako, Jpn) MAEDA M (Riken (inst. Of Physical And Chemical Res.), Wako, Jpn)
Journal Title;Anal Sci
Journal Code:G0673B
ISSN:0910-6340
VOL.21;NO.1;PAGE.25-29(2005)
Figure&Table&Reference;FIG.6, TBL.2, REF.21
Pub. Country;Japan
Language;English
Abstract;We developed a weak-affinity separation system for single-nucleotide polymorphisms (SNPs) based on capillary electrophoresis. In this approach, single-stranded DNA (ssDNA)-polyacrylamide (polyAAm) conjugate was used as a pseudo-immobilized affinity ligand to separate the target DNA, cytochrome P450 2C9 (CYP2C9), and its point mutant. The ligand DNA was designed to be complementary to the normal DNA, and the target DNA was electrophoretically separated by the difference in the affinity with the pseudo-immobilized ligand in the capillary. We showed that the separation efficiency was closely associated with the Tm value of double-stranded DNA (dsDNA) consisting of the target and ligand DNA, which depends on the measurement conditions, such as the base number of the ligand DNA and the concentration of Mg2+ in the buffer solution. (author abst.)
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