GSM5692686: pass2; Neisseria musculi; Tn-Seq
Lysis in TES buffer (50 mM Tris pH 8, 20 mM EDTA, 50 mM NaCl) with 1% SDS, Rnase A treatment followed by 25:24:1 Phenol chloroform isoamyl alcohol extraction with ethanol precipitation. DNA was sheared to 500 bp. Tn5 junction enrichment was performed on samples post end repair, A tailing and ligation to custom adaptors via NEBnext DNA kit. Amplicons from 100-600 bp were size selected and indexed by Nextera V2 dual index Kit. Samples were sequenced on an Illumina MiSeq as 75 bp paired end samples