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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.

SUBMITTER: Pack SP

PROVIDER: S-EPMC2034461 | biostudies-literature | 2007

REPOSITORIES: biostudies-literature

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Direct immobilization of DNA oligomers onto the amine-functionalized glass surface for DNA microarray fabrication through the activation-free reaction of oxanine.

Pack Seung Pil SP Kamisetty Nagendra Kumar NK Nonogawa Mitsuru M Devarayapalli Kamakshaiah Charyulu KC Ohtani Kairi K Yamada Kazunari K Yoshida Yasuko Y Kodaki Tsutomu T Makino Keisuke K

Nucleic acids research 20070821 17

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, particula ...[more]

PMID: 17715142

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Project description:Poly(ethylene terephthalate) (PET) and nylon find their main applications in working clothes, domestic furniture and as indoor decoration (curtains and carpets). The increasing attention on healthy lifestyle, together with protection and safety, gained a strong interest in today's society. In this context, reducing the flammability of textiles has been tackled by designing flame retardants (FRs) able to suppress or delay the flame propagation. Commercially available FRs for textiles often consist of brominated, chlorinated and organo-phosphorus compounds, which are considered a great concern for human health and for the environment. In this study, Deoxyribose Nucleic Acid (DNA) was investigated as a green and eco-friendly alternative to halogen-containing FRs. DNA is in fact able to provide flame retardant properties due to its intrinsically intumescent building blocks (deoxyribose, phosphoric-polyphosphoric acid, and nitrogen-containing bases). In a first step, anchor groups (i.e., carboxyl groups) for subsequent coupling of DNA were introduced to PET and nylon-6 fabrics via limited surface hydrolysis with Humicola insolens cutinase (HiC). Released monomer/oligomers were measured via HPLC (1 mM of BHET for PET and 0.07 mM of caprolactam from nylon after 72 h). In a next step, DNA immobilization on the activated polymers was studied by using three different coupling systems, namely: EDC/NHS, dopamine, and tyrosine. DNA coupling was confirmed via FT-IR that showed typical bands at 1,220, 970, and 840 cm-1. The tyrosine/DNA coupling on nylon fabrics resulted to be the most effective as certified by the lowest burning rate and total burning time (35 s, 150 mm, and 4.3 mm*s-1 for the blank and 3.5 s, 17.5 mm, and 5 mm* s-1 for nylon/tyrosine/DNA) which was also confirmed by FT-IR and ESEM/EDS measurements. Thermogravimetric analysis (TGA) further confirmed that tyrosine/DNA coated nylon showed a lower thermal degradation between 450 and 625°C when compared to the untreated samples.

Dataset Information

Direct immobilization of DNA oligomers onto the amine-functionalized glass surface for DNA microarray fabrication through the activation-free reaction of oxanine.

Publications

Direct immobilization of DNA oligomers onto the amine-functionalized glass surface for DNA microarray fabrication through the activation-free reaction of oxanine.

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