title: NGS SYK-6 RB-TnSeq library: parent TnSeq
design: 2 x 1 mL cell pellets were used to extract and purify gDNA, which underwent enzymatic fragmentation, end repair, and A-tailing with the NEBNext Ultra II FS Library Prep Kit (New England Biolabs, USA). Illumina Y-adapters were ligated onto gDNA templates and DNA was cleaned up with NEBNext Sample Purification Beads (New England Biolabs, USA). Specific primers were used to amplify the transposon-associated barcode region with Q5 High-Fidelity 2X Master Mix (New England Biolabs, USA). Samples were pooled and sent to Genewiz for sequencing.