Microarray fabrication with covalent attachment of DNA using bubble jet technology.
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.
Okamoto
T
T
Canon Research Center, 5-1, Morinosato-Wakamiya, Atsugi-shi, Kanagawa, 243-0193, Japan.
Suzuki
T
T
Yamamoto
N
N
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D017931:DNA Primers; D004867:Equipment Design; D005091:Exons; D016158:Genes, p53; D006801:Humans; D007438:Introns; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*; D015345:Oligonucleotide Probes; D017354:Point Mutation; D013873:Thionucleotides
D017931:DNA Primers; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D013873:Thionucleotides; D004247:DNA