fabrication of unimolecular double-stranded dna microarrays on solid surfaces for probing dna-proteindrug interactions制造的单分子的双链dna微阵列探测固体表面dna-proteindrug交互.pdfVIP

Molecules 2003, 8, 153- 168 molecules ISSN 1420-3049 Fabrication of Unimolecular Double -stranded DNA Microarrays on Solid Surfaces for Probing DNA-Protein/Drug Interactions Jinke Wang, Tongxiang Li, Yunfei Bai, Yi Zhu and Zuhong Lu* Chien-Shiung Wu Laboratory, Southeast University, Nanjing 210096, P.R. China * Author to whom correspondence should be addressed; E-mail: zhlu@ Abstract: We present a novel method for fabricating unimole cular double-stranded DNA microarrays on solid surfaces, which were used to probe sequence-specific DNA/protein interactions. For manufacturing the unimolecular double -stranded DNA microarrays, two kinds of special single-stranded oligonucleotides, constant oligonucleotide and target oligonucleotide, were chemically synthesized. The constant oligonucleotides with internal aminated dT were used to capture and immobilize the target oligonucleotides onto the solid surface, and also to provide a primer for later enzymatic extension reactions, while target oligonucleotides took the role of harbouring DNA-binding sites of DNA-binding proteins. The variant target oligonucleotides were annealed and ligated with the constant oligonucleotides to form the new unimolecular oligonucleotides for microspotting. The prepared unimolecular oligonucleotides were microspotted on aldehyde-derivatized glass slides to make partial-dsDNA microarrays. Finally, the partial-dsDNA microarrays were converted into a unimolecular complete-dsDNA microarray