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The separation of oligodeoxynucleotides having a single-base difference by affinity capillary electrophoresis using
Takahisa Anada1, Takumi Arisawa, Yoshihisa Ozaki
1Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka, Japan.
Abstract:
We have developed an affinity capillary electrophoresis for gene mutation assay. We show a method for separating a mixture of oligodeoxynucleotides having single-base difference using oligodeoxynucleotide-polyacrylamide conjugate. Because the conjugate behaved similarly to nonionic polyacrylamide in terms of migration rate, it can be used as a pseudoimmobilized affinity ligand in a polyacrylamide-coated capillary. Oligodeoxynucleotide having an oncogene sequence and a version with one base substituted were completely separated by this method. The magnesium-ion concentration was found to be a key factor in achieving efficient separation of oligodeoxynucleotides with the same chain length.