RNA sequencing of BCG Tokyo

BCG Tokyo 172 was cultured in Middlebrook 7H9-ADC-polysorbate 80 medium for 7 days (as log phase) or 14 days (as early stationary phase). RNA was extracted by using Direct-zol RNA MiniPrep Plus kit (Zymo Research, Irvine, CA). Quality of extracted RNA was assessed with PCR and TapeStation (Agilent). We determined the threshold of RNA integrity (RIN) as above 7 and rRNA ratio as above 0.8. Passed samples were analyzed in next generation sequencing service provided by Macrogen Japan.