Microarray fabrication with covalent attachment of DNA using Bubble Jet technology
Microarray fabrication with covalent attachment of DNA using Bubble Jet technology
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DOI：
10.1038/74507
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发表时间：
2000-04-01
影响因子：
46.9
通讯作者：
Yamamoto, N
中科院分区：
文献类型：
--
作者：
Okamoto, T;Suzuki, T;Yamamoto, N
我们已经开发了一种制造DNA微阵列的方法，该方法使用气泡喷射喷墨装置将5'端硫代寡核苷酸喷射到玻璃表面。通过与底物上的氨基和寡核苷酸探针上的巯基反应的杂双功能交联剂，将寡核苷酸共价连接到玻璃表面。使用这种方法，我们制作了携带64组18-mer寡核苷酸的DNA微阵列，编码p53肿瘤抑制基因突变“热点”中所有可能的三个碱基突变。用荧光标记合成的p53基因18聚寡核苷酸或从两种人口腔鳞状细胞癌（SCC）细胞系的基因组DNA中PCR扩增的p53基因片段进行筛选。这使我们能够区分匹配的杂种和1 bp不匹配的杂种。
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.