title: RNA sequencing results of wild-type TmaR (WT) and cells deleted for the tmaR gene (TmaR-KO)
design: RNA sequencing results of wild-type TmaR (WT) and cells deleted for the tmaR gene (TmaR-KO)
library_construction_protocol: Three independent cultures of wild type (WT) and ΔtmaR (TmaR-KO) were diluted 1:100 in fresh LB and allowed to grow until mid-logarithmic phase. Cells were harvested by centrifugation at 10,000 RPM and stored at -20 oC. Three independent cultures of wild type (WT) and ΔtmaR (TmaR-KO) were diluted 1:100 in fresh LB and allowed to grow until mid-logarithmic phase. Cells were harvested by centrifugation at 10,000 RPM and stored at -20 oC. RNA was extracted by Direct-zol RNA Miniprep Kit. Libraries were generated based on the RNAtag-Seq approach 20, with minor modifications. Briefly, 500 ng of triplicate RNA samples were fragmented at 94°C for 1.5 min, depleted of DNA, dephosphorylated, purified using RNAClean XP beads, and 3′ barcoded adapters were ligated to RNA samples. Barcoded RNAs were pooled, and Ribo-pool was used  to deplete rRNA. The samples were then subjected to template switching. The libraries were PCR amplified for sequencing and cleaned to cutoff fragments smaller than 500 bp.