A method for fabricating uni-dsDNA microarray chip for analyzing DNA-binding proteins.
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 5' end were firstly annealed to a same universal oligonucleotide with amino group at 5' 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 3' 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.
Wang
Jin K
JK
Chien-Shiung Wu Laboratory, Southeast University, Nanjing 210096, PR China. wangjinke@seu.edu.cn
Li
Tong X
TX
Lu
Zu H
ZH
D003362:Cost-Benefit Analysis; D004268:DNA-Binding Proteins / Q000032:analysis*; D008722:Methods; D046228:Microarray Analysis / Q000295:instrumentation*; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry; D016133:Polymerase Chain Reaction; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*
D004268:DNA-Binding Proteins; D009841:Oligonucleotides