Shalon D, Smith S J, Brown P O
Howard Hughes Medical Institute and Department of Biochemistry, Stanford University, California 94305, USA.
Genome Res. 1996 Jul;6(7):639-45. doi: 10.1101/gr.6.7.639.
Detecting and determining the relative abundance of diverse individual sequences in complex DNA samples is a recurring experimental challenge in analyzing genomes. We describe a general experimental approach to this problem, using microscopic arrays of DNA fragments on glass substrates for differential hybridization analysis of fluorescently labeled DNA samples. To test the system, 864 physically mapped lambda clones of yeast genomic DNA, together representing >75% of the yeast genome, were arranged into 1.8-cm x 1.8-cm arrays, each containing a total of 1744 elements. The microarrays were characterized by simultaneous hybridization of two different sets of isolated yeast chromosomes labeled with two different fluorophores. A laser fluorescent scanner was used to detect the hybridization signals from the two fluorophores. The results demonstrate the utility of DNA microarrays in the analysis of complex DNA samples. This system should find numerous applications in genome-wide genetic mapping, physical mapping, and gene expression studies.
在分析基因组时，检测并确定复杂DNA样本中各种不同个体序列的相对丰度是一个反复出现的实验挑战。我们描述了一种针对此问题的通用实验方法，即使用玻璃基板上的DNA片段微观阵列对荧光标记的DNA样本进行差异杂交分析。为了测试该系统，将代表酵母基因组75%以上的864个经物理定位的酵母基因组DNA λ克隆排列成1.8厘米×1.8厘米的阵列，每个阵列总共包含1744个元件。通过用两种不同荧光团标记的两组不同的分离酵母染色体同时杂交来表征这些微阵列。使用激光荧光扫描仪检测来自两种荧光团的杂交信号。结果证明了DNA微阵列在复杂DNA样本分析中的实用性。该系统应在全基因组遗传图谱绘制、物理图谱绘制和基因表达研究中有众多应用。
