In Vitro Selection of DNA Aptamers on Chips Using a Method for Generating Point Mutations

Abstract

We successfully developed a novel selection method for the acquisition of DNA aptamers that selectively recognize resorufin using on‐chip selection in combination with method for point mutations. This method proved efficient for selection for DNA aptamers of single‐stranded oligo‐DNAs. A genetic algorithm was applied to produce oligonucleotides for the combinatorial library. A fluorescent molecule, resorufin, was applied for the ligand selection as a target. The binding affinity of the library was analyzed by the DNA chip. This selection method of DNA ligands includes on‐chip selection and point‐mutated sequence, which the highest affinity was selected. The fluorescence intensity of the library on the DNA chip increased after three repetitions of the selection round. The average of response for the affinity test increased with each generation.

Keywords:

• Aptamer bioinformatics
• Genetic algorithm
• Lab‐on‐a‐chip
• Microarray

Acknowledgments

We would like to thank CombiMatrix Corp. and CombiMatrix KK for technical assistance.