title: RNA-seq of wild-type H37Rv and ΔsubI strains of Mycobacterium tuberculosis
design: RNA-seq of wild-type H37Rv and ΔsubI strains of Mycobacterium tuberculosis
library_name: SubI_KO_3_s
library_construction_protocol: WT and ΔsubI H37Rv were grown in triplicates in 7H9 medium supplemented with 10% ADC, 0,05% Tyloxapol, 1mM MgSO4, and 1mM cysteine until OD600 =0.8.   Total RNA was extracted from 10 mL of bacterial cultures grown to logarithmic phase (OD600nm = 0.8) using RNeasy kit (Qiagen).  Total Bulk RNA sequencing was performed Each extracted total RNA sample was barcoded with CEL-seq primers during the enzymatic lysis. It was then followed by fragmentation, end repair, poly(A)-tailing, reverse transcription, and second-strand synthesis. After second-strand synthesis, the barcoded samples were pooled into one library and amplified using in vitro transcription. Further, ribosomal aRNA was depleted. After depletion, adapter ligation, reverse transcription, and indexing PCR were performed to prepare a cDNA library.  