Nucleic Acids Research Advance Access originally published online on August 21, 2007
Nucleic Acids Research 2007 35(17):e110; doi:10.1093/nar/gkm619
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Nucleic Acids Research, 2007, Vol. 35, No. 17 e110
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Methods Online |
Direct immobilization of DNA oligomers onto the amine-functionalized glass surface for DNA microarray fabrication through the activation-free reaction of oxanine
1Institute of Advanced Energy, Kyoto University, 2CREST, JST (Japan Science and Technology Agency), Kyoto University, Gokasho, Uji 611-0011, 3NGK Insulators, Ltd, GENESHOT project, Mizuho, Nagoya 467-8530 and 4Kyoto Nanotechnology Cluster, Kyoto University, Gokasho, Uji 611-0011, Japan
*To whom correspondence should be addressed. Tel: +81 774 38 3517; Fax: +81 774 38 3524; Email: kmak{at}iae.kyoto-u.ac.jp
Received March 7, 2007. Revised July 18, 2007. Accepted July 30, 2007.
Oxanine having an O-acylisourea structure was explored to see if its reactivity with amino group is useful in DNA microarray fabrication. By the chemical synthesis, a nucleotide unit of oxanine (Oxa-N) was incorporated into the 5'-end of probe DNA with or without the -(CH2)n- spacers (n = 3 and 12) and found to immobilize the probe DNA covalently onto the NH2-functionalized glass slide by one-pot reaction, producing the high efficiency of the target hybridization. The methylene spacer, particularly the longer one, generated higher efficiency of the target recognition although there was little effect on the amount of the immobilized DNA oligomers. The post-spotting treatment was also carried out under the mild conditions (at 25 or 42°C) and the efficiencies of the immobilization and the target recognition were evaluated similarly, and analogous trends were obtained. It has also been determined under the mild conditions that the humidity and time of the post-spotting treatment, pH of the spotting solution and the synergistic effects with UV-irradiation largely contribute to the desired immobilization and resulting target recognition. Immobilization of DNA oligomer by use of Oxa-N on the NH2-functionalized surface without any activation step would be employed as one of the advanced methods for generating DNA-conjugated solid surface.