title: GSM8577339: Control population, 45 hr; Escherichia coli; Tn-Seq
library_construction_protocol: Genomic DNA was extracted from both control and enriched populations using a GeneAll Exgene™ Cell SV kit (GeneAll Biotechnology, Seoul, Korea). The extracted gDNA was digested with the restriction enzyme MmeI. The digested DNA fragments, which include a short adjacent region of the Himar1 transposon insertion site, were ligated to an adapter with an NN tail. The adapter-ligated DNAs were used as a template for amplification, with the Tn-IR-P7 primer (which primes the transposon inverted repeat and includes an overhang for the P7 sequence) and the Adapter-P5 primer (which primes the adaptor and includes the P5 sequence). This produced the Tn-seq libraries for Illumina sequencing. The constructed libraries were validated using X-Mark LabChip with the DNA NGS 3K reagent kit (PerkinElmer, Waltham, MA, USA). The concentration of the libraries was measured using Qubit™ dsDNA Quantification Assay Kit (Thermo, Waltham, MA, USA).