Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10824940 | Journal of Biochemical and Biophysical Methods | 2005 | 11 Pages |
Abstract
This paper describes an approach for preparing unimolecular double-stranded DNA (uni-dsDNA) microarray chip. In this method, the various target oligonucleotides containing a reverse complementary sequence at 5â² end were firstly annealed to a same universal oligonucleotide with amino group at 5â² end and immobilized on aldehyde-derivatized glass slide. An on-chip DNA polymerization reaction was then performed to elongate the universal oligonucleotides. After a denaturation and a followed intra-strand annealing, a hairpin structure was formed at the free 3â² end of the immobilized oligonucleotides. Finally, another on-chip DNA polymerization was done to synthesize the uni-dsDNA microarray. Combining with a PCR amplification of chemically synthesized target oligonucleotides, this method was much cost-effective for production of the uni-dsDNA microarray. The uni-dsDNA microarray was verified applicable for detecting the presence and monitoring the DNA-binding activity of the sequence-specific DNA-binding proteins.
Keywords
Nonidet P40NP-40AP1PBSdNTPSDSSSCHEPESDTTNF-κBBSADNAN-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acidbovine serum albuminEDTAEthylenediaminetetraacetic aciddeoxyribonucleic acidtumor necrosis factor αdeoxynucleoside triphosphatedithiothreitolFabricationsodium dodecyl sulfateTNF-αnuclear factor κBPhosphate-buffered salinepolymerase chain reactionPCRactivator protein 1DNA-binding proteinhigh-performance liquid chromatographyHPLC
Related Topics
Life Sciences
Biochemistry, Genetics and Molecular Biology
Biochemistry
Authors
Jin K. Wang, Tong X. Li, Zu H. Lu,