Broad Genome Sequencing of Environmental and Clinical Strains and Genotyping

Abstract

The Next-Generation Sequencing revolution had a great impact on the genomics of Pseudomonas aeruginosa. Since the first release of the P. aeruginosa PAO1 genome, there are more than 5700 genomes published. This wealth of information has been accompanied by the development of bioinformatic tools for handling genomic and phenotypic data. Bioinformatics, indeed, become de facto a big data science. In this chapter, we give a brief historical overview of the knowledge gained from P. aeruginosa genome sequencing, then we describe the wet-lab procedure to extract the DNA and prepare the library for broad genome sequencing using Illumina MiSeq technology. As last, we describe three user-friendly bioinformatics procedures to infer the P. aeruginosa genotype, starting from NGS data, with the Multi-Locus Sequence Typing method, and visualize it as a minimum spanning tree.

Keywords: DNA extraction; Genome sequencing; Genotyping; Minimum spanning tree; Next-generation sequencing; Pseudomonas aeruginosa; eBURST.