GSM9681820: C068_B_BAL_IGRAneg; Homo sapiens; RNA-Seq
Cell suspensions containing 0.5 x 106 live cells were pelleted by centrifuging at 400 x G for 5 minutes at 4oC and pellets suspended in TRI reagent (Sigma-Aldrich). Samples were vortexed at high speed for around 20 seconds before storage at -80oC until RNA extraction. RNA extraction was performed using the DirectZol RNA Microprep kit (Zymo Technologies), as per manufacturer's instructions, including on-column DNase I digestion. Extracted RNA from whole BAL cell pellets was subjected to Agilent TapeStation quality control analysis and RNA integrity (RIN) values determine to be ≥7.4 and ≤9.9, except for one sample (T219_1, from active TB patient #219), which had a RIN value of 4.1. 200 ng of total RNA was used for library preparation using the RNA library prep kit with Polaris ribosomal RNA depletion (Watchmaker Genomics). Approximately 2.5 x107 100 base pair paired end reads were obtained per sample using an Illumina NovaSeq X sequencer.