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A fabrication method of linkers to covalently anchor nucleic acid probes (such as, oligonucleotides, PCR-products or peptide nucleic acid oligomers) on glass was developed by alternatively covalently assembling the different molecular components. The linkers with controllable length, flexibility and hydrophilicity could be prepared simply by dipping subtracts alternatively in different chemical solutions. A dialkoxy aminosilane (N-(2-aminoethyl)-3-aminopropylmethyl dimethoxylsilane, AEAPS) was chosen to substitute common used trialkoxy aminosilane to modify glass surface. The end groups of the linkers were formed as aldehyde or amino group, which were successfully used for attaching prefabricated DNA or in situ synthesis of oligonucleotides, respectively. The both experiments showed that the linker produce good reproducibility and uniformity of fluorescent hybridizing images, which can distinguish an internal single base mismatch in 20mer oligonucleotides

Assembly method fabricating linkers for covalently bonding DNA on glass surface