Library on a slide for bacterial comparative genomics
1 Department of Epidemiology, University of Michigan School of Public Health, 109 Observatory Street, Ann Arbor, MI48109, USA
2 Department of Biostatistics, University of Michigan School of Public Health, 109 Observatory Street, Ann Arbor, MI48109, USA
3 Department of Pediatric Infectious Disease, University of Michigan Medical School, 1521 Simpson, Ann Arbor, MI 48109, USA
BMC Microbiology 2004, 4:12 doi:10.1186/1471-2180-4-12Published: 22 March 2004
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.