DNA microarray fabricated on poly(acrylic acid) brushes-coated porous silicon by in situ rolling circle amplification.
Microarrays hold considerable promise in large-scale biology on account of their analytical, massive and parallel nature. In a step toward further enabling such a capability, we describe the application of rolling circle amplification (RCA) for a sensitive and multiplex detection of nucleic acid targets on oligonucleotide-conjugated polymer brushes covalently grown from porous silicon. Both RCA and polymer brushes have been taken to increase the loading quantity of target molecules and thus improve the detection sensitivity without loss of multiplexing. Besides, polymer brushes were employed to protect porous silicon and to provide biologically simulated environments, making the attached biomolecules maintain bioactivity. This approach can reach a detection limit of 0.1 nM target analytes and three orders of magnitude dynamic range of 0.1-100 nM, with a fluorescence scanner. A two-colour DNA microarray was achieved via RCA of two kinds of circular DNA targets on one chip simultaneously. The porous silicon chip-based RCA technique is promising for the multiplex detection of deoxynucleic acids on microarrays.
State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, Jiangsu, PR China.
Cuie
C
Wang
Xue-Mei
XM
Jia
Chuan
C
Jiang
Guang-Nan
GN
Zhuang
Qin
Q
Yan
Shou-Jun
SJ
Xiao
D000180:Acrylic Resins / Q000737:chemistry*; D004247:DNA / Q000032:analysis*; D021141:Nucleic Acid Amplification Techniques; D020411:Oligonucleotide Array Sequence Analysis; D016062:Porosity; D012825:Silicon / Q000737:chemistry*
D000180:Acrylic Resins; C006903:carbopol 940; D004247:DNA; D012825:Silicon