Transposon insertion site sequencing (TIS) of Pseudomonas aeruginosa

Transposon insertion site sequencing (TIS) is a technique that determines the insertion profile of a transposon mutant library by massive parallel sequencing of transposon-genomic DNA junctions. Because the transposon insertion profile reflects the abundance of each mutant in the library, it provides information to assess the fitness contribution of each genetic locus of a bacterial genome in a specific growth condition or strain background. Although introduced only about a dozen years ago, TIS has become an important tool in bacterial genetics that provides clues to study biological functions and regulatory mechanisms. Here, I describe a protocol for generating high density transposon insertion mutant libraries and preparing Illumina sequencing samples for mapping the transposon junctions of the transposon mutant libraries using Pseudomonas aeruginosa as an example.

Automated summary

Transposon insertion site sequencing (TIS) is a powerful technique for determining the insertion profile of a transposon mutant library, providing insights into the fitness contributions of genetic loci in bacterial genomes under specific growth conditions. Despite being relatively new, TIS has emerged as a crucial tool in bacterial genetics research.