4.6 Article

Library on a slide for bacterial comparative genomics

Journal

BMC MICROBIOLOGY
Volume 4, Issue -, Pages -

Publisher

BMC
DOI: 10.1186/1471-2180-4-12

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Funding

  1. NIAID NIH HHS [AI51675, R03 AI054406, AI054406, R01 AI051675] Funding Source: Medline
  2. NIDCD NIH HHS [R01 DC005840, DC005840] Funding Source: Medline
  3. NIDDK NIH HHS [DK 055496, R01 DK055496] Funding Source: Medline

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Background: 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. Results: 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 noncompeting hybridization strategy. Results from the array hybridization matched perfectly with previous tests. Conclusions: 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.

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