Zhang Lixin, Srinivasan Usha, Marrs Carl F, Ghosh Debashis, Gilsdorf Janet R, Foxman Betsy
Department of Epidemiology, University of Michigan School of Public Health, 109 Observatory Street, Ann Arbor, MI48109, USA.
BMC Microbiol. 2004 Mar 22;4:12. doi: 10.1186/1471-2180-4-12.
We describe a novel application of microarray technology for comparative genomics of bacteria in which libraries of entire genomes rather than the sequence of a single genome or sets of genes are arrayed on the slide and then probed for the presence or absence of specific genes and/or gene alleles.
We first adopted a 96-well high throughput working protocol to efficiently isolate high quality genomic DNA. We then optimized conditions to print genomic DNA onto a glass slide with high density (up to 15000 spots) and to sensitively detect gene targets in each genome spot using fluorescently labeled DNA probe. Finally, we created an E. coli reference collection array and probed it for the presence or absence of the hemolysin (hly) gene using a dual channel non-competing hybridization strategy. Results from the array hybridization matched perfectly with previous tests.
This new form of microarray technology, Library on a Slide, is an efficient way for sharing and utilizing large strain collections in comparative genomic analyses.
我们描述了一种微阵列技术在细菌比较基因组学中的新应用，即将整个基因组文库而非单个基因组或基因集的序列排列在载玻片上，然后检测特定基因和/或基因等位基因的有无。
我们首先采用96孔高通量工作方案来高效分离高质量基因组DNA。然后优化条件，将基因组DNA高密度（多达15000个点）打印到载玻片上，并使用荧光标记的DNA探针灵敏地检测每个基因组点中的基因靶标。最后，我们创建了一个大肠杆菌参考集合阵列，并使用双通道非竞争性杂交策略检测其中溶血素（hly）基因的有无。阵列杂交结果与先前的测试完美匹配。
这种新形式的微阵列技术——载玻片上的文库，是在比较基因组分析中共享和利用大量菌株集合的有效方法。
