TnSeq of N.aromaticivorans
DNA was extracted from transposon mutagenized library of N.aromaticivorans cultured on Earth using Qiagen Maxi Prep Kit and purified using Zymo ZR-96 DNA Cleanup and Concentrator-5 kit. Nextera libraries were prepared for each sample following the Illumina Nextera DNA Flex Library Prep protocol. Libraries were quantified using an Agilent 4200 TapeStation and diluted to 1 nM as a template for transposable element (TE) library construction via Nested PCR. Multiplex PCR reactions were then performed using primers specific to the TE and the index i5 primer. The final libraries were analyzed using the Agilent 4200 TapeStation, purified with 0.8X AMPure XP beads, washed with 80% ethanol twice, and eluted with elution buffer. The libraries were loaded on NextSeq 500/550 High Output (75 cycle) v2.5 flow cells configured to generate 75bp paired-end reads. The demultiplexing of the reads was performed using bcl2fastq version 2.20.0.