GSM9130479: After selection replicate 2; Escherichia coli str. K-12 substr. MG1655; Tn-Seq
Genomic DNA was extracted as per the NEB Genomic DNA preparation kit (T3010) for gram negative bacteria. Genomic DNA was sheared using Covaris ML230 into ~1 kb fragments. End repaired using NEB end repair kit (E6050). Poly-C tailed using NEB terminal transferase (M0315) and dCTP. Purified with sample purification beads and region amplified using primers specific to the transposon paired with a poly-G primer that introduces a known sequence to the other end. A second, nested PCR was done to introduce illumina barcode handles.