title: Illumina HiSeq 2000 paired end sequencing; Global mRNA transcriptome analysis of wild type and Hfq mutant strains of Serratia marcescens by RNAseq
design: Global mRNA transcriptome analysis of wild type and Hfq mutant strains of Serratia marcescens by RNAseq
library_construction_protocol: Bacterial cultures, four independent cultures for each strain, were grown in liquid LB media (10 g/l tryptone, 5 g/l yeast extract, 5 g/l NaCl) with good aeration at 30C to mid log phase (5 h) RNA was isolated from bacterial cells by hot phenol extraction. Contaminating gDNA was removed from using the DNA-free Kit (Ambion), followed by ethanol precipitation of the RNA. Ribosomal RNA (rRNA) was depleted using the Ribo-zero rRNA Removal Kit for Gram-negative bacteria (Epicentre). Depleted RNA samples were purified by ethanol precipitation and resuspension in RNase free water, then subsequently purified with AMPure XP beads (Agencourt) to remove RNA fragments < 200bp. Illumina TruSeq
attributes.tag: Experimental Factor: none
attributes.value: genetic modification
attributes.tag: Experimental Factor: wild type genotype
attributes.value: genotype