title: Direct immobilization of DNA oligomers onto the amine-functionalized glass surface for DNA microarray fabrication through the activation-free reaction of oxanine.
abstract: 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 degrees 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.
authors: Institute of Advanced Energy, Kyoto University, Japan.
authors: Seung Pil
authors: SP
authors: Pack
authors: Nagendra Kumar
authors: NK
authors: Kamisetty
authors: Mitsuru
authors: M
authors: Nonogawa
authors: Kamakshaiah Charyulu
authors: KC
authors: Devarayapalli
authors: Kairi
authors: K
authors: Ohtani
authors: Kazunari
authors: K
authors: Yamada
authors: Yasuko
authors: Y
authors: Yoshida
authors: Tsutomu
authors: T
authors: Kodaki
authors: Keisuke
authors: K
authors: Makino
mesh_terms: D000588:Amines / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry*; D005898:Glass; D006813:Humidity; D006863:Hydrogen-Ion Concentration; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry; D011684:Purine Nucleosides / Q000737:chemistry*; D013696:Temperature; D013997:Time Factors
chemical_list: D000588:Amines; D015342:DNA Probes; D015345:Oligonucleotide Probes; D011684:Purine Nucleosides; C468766:oxanine
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