Identification of Invasion-Related Genes in P. aeruginosa

The detection of invasion-related genes belonging to the T1SS, T2SS and T3SS was performed via PCR in a Veriti thermal cycler (Applied Biosystem, Foster City, CA, USA) according to the protocol described by Rodrigues et al. [32 (link)]. Visualization of PCR products was performed via 1.5% agarose gel electrophoresis at 110 V for 45 min in TAE 1× buffer (89 nM Tris-borate and 2 mM EDTA pH 8.0). As molecular weight marker, 1 Kb DNA ladder (Invitrogen™) was used, gel stained with SyberSafe (Invitrogen™, Carlsbad, CA, USA)) and differentiation of bands visualized under ultraviolet light. In addition, the pigment production and mucoid phenotype of P. aeruginosa isolates were verified by observing bacterial growth on MHA agar plates and slants.