title: GSM7147233: H37Rv, 15min unexposed, rep1; Mycobacterium tuberculosis; RNA-Seq
library_construction_protocol: Bacterial pellets were used for RNA extraction using the Quick-RNA Fungal/Bacterial Miniprep kit (Zymo Research). Briefly, the pellet was resuspended in Lysis buffer and transferred to a ZR BashingBead Lysis tube. A bead beating procedure was performed in a BioSpec Mini-Beadbeater-24 (6 cycles of 45 s at 3,800 rpm, with 3 min intervals on a cooler rack). RNA in the supernatant was isolated using Zymo-spin columns, with an in-column DNAse I treatment, and eluted in nuclease-free water. A second DNAse treatment was performed on the isolated RNA using the TURBO DNAse reagent (Thermo Fisher Scientific), and RNA was further purified using the RNA Clean & Concentrator kit (Zymo Research). A total RNA stranded library was prepared using the Zymo-Seq RiboFree Total RNA Library Kit (Zymo Research). Briefly, 500 ng of RNA were used as input to construct the library, which included a rRNA depletion step of 30 min followed by 14 cycles of PCR. 75bp paired-end (PE) sequencing was conducted using a NextSeq 500 Mid Output Illumina platform.