Lashkari D A, DeRisi J L, McCusker J H, Namath A F, Gentile C, Hwang S Y, Brown P O, Davis R W
Department of Genetics, Stanford University, CA 94305, USA.
Proc Natl Acad Sci U S A. 1997 Nov 25;94(24):13057-62. doi: 10.1073/pnas.94.24.13057.
We have developed high-density DNA microarrays of yeast ORFs. These microarrays can monitor hybridization to ORFs for applications such as quantitative differential gene expression analysis and screening for sequence polymorphisms. Automated scripts retrieved sequence information from public databases to locate predicted ORFs and select appropriate primers for amplification. The primers were used to amplify yeast ORFs in 96-well plates, and the resulting products were arrayed using an automated micro arraying device. Arrays containing up to 2,479 yeast ORFs were printed on a single slide. The hybridization of fluorescently labeled samples to the array were detected and quantitated with a laser confocal scanning microscope. Applications of the microarrays are shown for genetic and gene expression analysis at the whole genome level.
我们已经开发出了酵母开放阅读框(ORF)的高密度DNA微阵列。这些微阵列可监测与开放阅读框的杂交情况,用于诸如定量差异基因表达分析和序列多态性筛选等应用。自动化脚本从公共数据库检索序列信息,以定位预测的开放阅读框并选择合适的引物进行扩增。这些引物用于在96孔板中扩增酵母开放阅读框,所得产物使用自动化微阵列设备进行排布。在一张载玻片上打印了包含多达2479个酵母开放阅读框的阵列。用激光共聚焦扫描显微镜检测并定量荧光标记样品与阵列的杂交情况。展示了这些微阵列在全基因组水平的遗传和基因表达分析中的应用。