Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNA was isolated, from blood from tuberculosis progressors and Treatment reponse cohorts, and IGRA-ve healthy controls simultaneously, from 1ml whole blood using the MagMAX™ for Stabilized Blood Tubes RNA Isolation Kit . Globin RNA was depleted from the total RNA (1.5–2 μg) using the human GLOBINclear kit. Samples (200 ng) with an RQS > 6 were used to prepare a cDNA library using the TruSeq Stranded mRNA HT Library Preparation Kit (Illumina). The tagged libraries were sized and quantitated in duplicate (Agilent TapeStation system) using D1000 ScreenTape and reagents (Agilent), normalised, pooled and then clustered using the HiSeq® 3000/4000 PE Cluster Kit (Illumina). The libraries were imaged and sequenced on an Illumina HiSeq 4000 sequencer using the HiSeq® 3000/4000 SBS kit (Illumina) at a minimum of 25 million paired-end reads (100 bp) per sample