This method is significant because it can be used in diverse bacterial species to identify essential genes, antibiotic resistance genes and genes important for in vivo fitness during infection. The main advantage of this technique is the mechanica
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We describe a method to generate saturating transposon mutant libraries in Gram-negative bacteria and subsequent preparation of DNA amplicon libraries for high-throughput sequencing. As an example, we focus on the ESKAPE pathogen, Acinetobacter baumannii, but this protocol is amenable to a wide range of Gram-negative organisms.