GSM3615064: RvB_M_Smeg; Mycolicibacterium smegmatis; RNA-Seq
Total RNA was isolated from the  sample by trizol method. The quality of the total RNA was checked on 1% formaldehyde denature Agarose gel and quantified using Qubit fluorometer. Isolated RNA from given sample was treated with MICROBExpress Bacterial mRNA Enrichment kit to enrich microbial mRNA from total RNA. The libraries were prepared using TruSeq Stranded RNA Library Preparation Kit and as per its described protocol. The fragmented mRNA was converted into first-strand cDNA, followed by second-strand cDNA generation, A-tailing, adapter ligation and finally ended by limited number of PCR amplification of the adaptor-ligated libraries. Library quantification and qualification was performed using DNA High Sensitivity Assay Kit.