Tn-seq inBordetella pertussis: input 5
A 2.5 hour digestion of 3mg of genomic DNA with 6U MmeI (New England Biolabs) was performed, and bands between 1-2kb were excised. Adapters were ligated onto the fragments overnight, and then the products were PCR purified to remove excess adapter. A PCR reaction was performed to amplify the Tn-genomic junction, add a barcode sequence, and add required Illumina sequences (P7, flow cell binding) (S8 Table). This sample was purified by size exclusion (about 125 bp).