A DNA microarray system for analyzing complex DNA samples using two-color fluorescent probe hybridization
A DNA microarray system for analyzing complex DNA samples using two-color fluorescent probe hybridization
复制标题
利用双色荧光探针杂交分析复杂 DNA 样品的 DNA 微阵列系统
DOI：
10.1101/gr.6.7.639
复制
发表时间：
1996-07-01
期刊：
影响因子：
7
通讯作者：
Brown, PO
中科院分区：
文献类型：
--
作者：
Shalon, D;Smith, SJ;Brown, PO
检测和确定复杂DNA样品中不同个体序列的相对丰度是分析基因组中反复出现的实验挑战。我们描述了一个一般的实验方法，这个问题，使用显微镜阵列的DNA片段的玻璃基板上的荧光标记的DNA样品的差异杂交分析。为了测试该系统，将864个酵母基因组DNA的物理映射λ克隆（一起代表> 75%的酵母基因组）排列成1.8 cm x 1.8 cm阵列，每个阵列总共包含1744个元件。微阵列的特点是同时杂交的两个不同的组分离的酵母染色体标记有两种不同的荧光团。激光荧光扫描仪用于检测来自两个荧光团的杂交信号。结果表明DNA微阵列在复杂DNA样品分析中的实用性。该系统将在全基因组遗传作图、物理作图和基因表达研究中得到广泛应用。
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.