NextSeq 550 sequencing; RNA-seq of Streptococcus pneumoniae serotype 6B; wild type strain and 2 different double mutants grown in Todd-Hewitt media supplemented with 0.5% yeast extract (THY)
RNA-seq of Streptococcus pneumoniae serotype 6B; wild type strain and 2 different double mutants grown in Todd-Hewitt media supplemented with 0.5% yeast extract (THY)
RNA for RNAseq were extracted from double mutant strains cultured to an OD600 0.4-0.5  Bacterial strains were cultured in Todd-Hewitt supplemented with  0.5% yeast extract (THY), to an OD600 0.4-0.5  This package was also used for differential gene expression. This analysis used a p-value cut-off of 0.01 and a cut-off of 1.5 log-fold-change to be considered significantly differentially expressed. RNA for RNAseq were extracted from double mutant strains cultured to an OD600 0.4-0.5 using Mirvana RNA kit (Applied biosystems) with an additional physical lysis step using 0.1 mm glass beads (MP Biomedicals), treated with Turbo DNAse (Applied biosystems), and deleted of ribosomal RNA using Ribo-Zero Magnetic Kit Bacteria (Illumina) before preparation of sequencing libraries  sequencing libraries were prepared using the KAPA RNA HyperPrep kit (Roche Diagnostics) and 8 cycles of amplificationKAPA RNA HyperPrep Kit - Automated Library Prep
Experimental Factor: strain
6B1_4