In vitro selection of DNA aptamers on chips using a method for generating point mutations

被引:20
|
作者
Asai, R [1 ]
Nishimura, SI [1 ]
Aita, T [1 ]
Takahashi, K [1 ]
机构
[1] AIST, Computat Biol Res Ctr, Koutou Ku, Tokyo 1350064, Japan
关键词
aptamer bioinformatics; genetic algorithm; lab-on-a-chip; microarray;
D O I
10.1081/AL-120029741
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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.
引用
收藏
页码:645 / 656
页数:12
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