Analytical Sciences

Oligodeoxynucleotide-Modified Capillary for Electrophoretic Separation of Single-Stranded DNAs with a Single-Base Difference

Takahisa ANADA,* Masako OGAWA,* Hisashi YOKOMIZO,* Yoshihisa OZAKI,* Tohru TAKARADA,* Yoshiki KATAYAMA,* and Mizuo MAEDA*^,** 

*Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 6-10-1, Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan
**Bioengineering Laboratory, RIKEN (The Institute of Physical and Chemical Research), 2-1, Hirosawa, Wako 351-0198, Japan

We describe here a method of affinity capillary electrophoresis in which oligodeoxynucleotide (ODN) was immobilized onto the inner surface of the capillary. The immobilized ODN functioned successfully as an affinity ligand for sequence-based DNA separation. Six- or 12-mer ODN with a sequence complementary to one of the c-K-ras gene was used as an immobilized ligand. When the 12-mer ODN was used, the detection peak for the complementary ODN disappeared selectively, while the single-base mutant was detected as usual. In contrast, when the 6-mer ODN was used as the affinity ligand with a mixture of the complementary ODN and its single-base mutant, it was possible to detect both as completely separate peaks. That is, the separation mode was dependent on the base number of the immobilized ODN used as an affinity ligand.

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