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Microarray fabrication with covalent attachment of DNA using Bubble Jet technology

Nature Biotechnology volume 18, pages 438441 (2000) | Download Citation

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Abstract

We have developed a method for fabricating DNA microarrays that uses a Bubble Jet ink jet device to eject 5′-terminal-thiolated oligonucleotides to a glass surface. The oligonucleotides are covalently attached to the glass surface by heterobifunctional crosslinkers that react with the amino group on the substrate and a thiol group on the oligonucleotide probe. Using this method, we fabricated DNA microarrays that carried 64 groups of 18-mer oligonucleotides encoding all possible three-base mutations in the mutational “hot spot” of the p53 tumor-suppressor gene. These were screened with a fluorescently labeled synthetic 18-mer oligonucleotide derived from the p53 gene, or segments of the p53 gene that had been PCR amplified from genomic DNA of two cell lines of human oral squamous cell carcinoma (SCC). This allowed us to discriminate between matched hybrids and 1 bp-mismatched hybrids.

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References

  1. 1.

    et al. Microarrays: biotechnology's discovery platform for functional genomics. Tibtech 16, 301– 306 (1998).

  2. 2.

    Massively parallel genomics. Science 277, 393–395 (1997).

  3. 3.

    , , & Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science 270, 467–470 ( 1995).

  4. 4.

    & Bubble jet recording. J. Inst. Image Electron. Eng. Jpn.. 11, 66– 71 (1982).

  5. 5.

    , & Bubble generation mechanism in the bubble jet recording process. J. Imaging Tech. 14, 120– 124 (1988).

  6. 6.

    , , , & Single nucleotide polymorphic discrimination by an electronic dot blot assay on semiconductor microchip. Nat. Biotechnol. 17, 365–370 ( 1999).

  7. 7.

    et al. Database of p53 gene somatic mutations in human tumors and cell lines: updated compilation and future prospects. Nucleic Acids Res. 25, 151–157 ( 1997).

  8. 8.

    , & Covalent attachment of synthetic DNA to self-assembled monolayer films. Nucleic Acids Res. 24, 3031–3039 (1996).

  9. 9.

    , , , & Preparation and characterization of hetero-bifunctional cross-linking reagents for protein modifications. Chem. Pharm. Bull. 29, 1130–1135 (1981).

  10. 10.

    & Most human squamous cell carcimomas in the oral cavity contain mutated p53 tumor-suppressor genes. Oncogene 7, 927–933 ( 1992).

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Acknowledgements

The authors express their sincere appreciation to Prof. Nobuo Tsuchida of Tokyo Medical and Dental University for kindly providing HSC4, HSC5, and the PCR primers, Drs. Fumie Hosoda and Hitoshi Ichikawa of National Cancer Research Center for invaluable advice.

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Affiliations

  1. Canon Research Center, 5-1, Morinosato-Wakamiya , Atsugi-shi, Kanagawa, 243-0193, Japan

    • Tadashi Okamoto
    • , Tomohiro Suzuki
    •  & Nobuko Yamamoto

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Correspondence to Nobuko Yamamoto.

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DOI

https://doi.org/10.1038/74507

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