LIGHT-GENERATED OLIGONUCLEOTIDE ARRAYS FOR RAPID DNA-SEQUENCE ANALYSIS
LIGHT-GENERATED OLIGONUCLEOTIDE ARRAYS FOR RAPID DNA-SEQUENCE ANALYSIS
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DOI：
10.1073/pnas.91.11.5022
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发表时间：
1994-05-24
影响因子：
11.1
通讯作者：
FODOR, SPA
中科院分区：
文献类型：
--
作者：
PEASE, AC;SOLAS, D;FODOR, SPA
在分子生物学的许多领域，都需要快速提取和分析遗传信息；然而，目前的 DNA 序列分析技术速度缓慢且劳动强度大。我们在此报告如何使用现代光刻技术通过生成密集堆积的寡核苷酸探针的小型化阵列来促进序列分析。这些探针阵列或 DNA 芯片随后可用于并行 DNA 杂交分析，直接产生序列信息。在初步实验中，通过 16 个化学反应循环产生了 256 种不同八核苷酸的 1.28 x 1.28 厘米阵列，需要 4 小时才能完成。然后通过落射荧光显微镜检测荧光标记的寡核苷酸靶标的杂交模式。互补探针的荧光信号比单碱基对或双碱基对杂交错配的荧光信号强5-35倍，证明了互补序列识别的特异性。这种方法应该被证明是人类遗传学和诊断、病原体检测和 DNA 分子识别快速研究的有力工具。
In many areas of molecular biology there is a need to rapidly extract and analyze genetic information; however, current technologies for DNA sequence analysis are slow and labor intensive. We report here how modern photolithographic techniques can be used to facilitate sequence analysis by generating miniaturized arrays of densely packed oligonucleotide probes. These probe arrays, or DNA chips, can then be applied to parallel DNA hybridization analysis, directly yielding sequence information. In a preliminary experiment, a 1.28 x 1.28 cm array of 256 different octanucleotides was produced in 16 chemical reaction cycles, requiring 4 hr to complete. The hybridization pattern of fluorescently labeled oligonucleotide targets was then detected by epifluorescence microscopy. The fluorescence signals from complementary probes were 5-35 times stronger than those with single or double base-pair hybridization mismatches, demonstrating specificity in the identification of complementary sequences. This method should prove to be a powerful tool for rapid investigations in human genetics and diagnostics, pathogen detection, and DNA molecular recognition.