Synergistic effects of epoxy- and amine-silanes on microarray DNA immobilization and hybridization.
Most microarray slides are manufactured or coated with a layer of poly(L-lysine) or with silanes with different chemical functional groups, for the attachment of nucleic acids on to their surfaces. The efficiency with which nucleic acids bind to these surfaces is not high, because they can be washed away, especially in the case of spotting oligonucleotides. In view of this, we have developed a method to increase the binding capacity and efficiency of hybridization of DNA on to derivatized glass surfaces. This makes use of the synergistic effect of two binding interactions between the nucleic acids and the coating chemicals on the surface of the glass slides. The enhanced binding allows the nucleic acids to be bound tightly and to survive stringency washes. When immobilized, DNA exhibits a higher propensity for hybridization on the surface than on slides with only one binding chemical. By varying the silane concentrations, we have shown that maximal DNA oligonucleotide binding on glass surfaces occurs when the percentage composition of both of the surface-coating chemicals falls to 0.2%, which is different from that on binding PCR products. This new mixture-combination approach for nucleic-acid binding allows signals from immobilization and hybridization to have higher signal-to-noise ratios than for other silane-coated methods.
Chiu
Sung-Kay
SK
Department of Research and Development, U-Vision Biotech Inc., 3F No.132 Lane 235 Pao-Chiao Rd, Hsin-Tien City 231, Taipei, Taiwan.
Hsu
Mandy
M
Ku
Wei-Chi
WC
Tu
Ching-Yu
CY
Tseng
Yu-Tien
YT
Lau
Wai-Kwan
WK
Yan
Rong-Yih
RY
Ma
Jing-Tyan
JT
Tzeng
Chi-Meng
CM
D000588:Amines / Q000737:chemistry*; D001665:Binding Sites; D004247:DNA / Q000737:chemistry*; D004852:Epoxy Compounds / Q000737:chemistry*; D005898:Glass; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009696:Nucleic Acids / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012821:Silanes / Q000737:chemistry*
D000588:Amines; D004852:Epoxy Compounds; D009696:Nucleic Acids; D012821:Silanes; D004247:DNA