title: Illumina HiSeq 1000 paired end sequencing; RNA-seq analysis of primary, processed and total RNAs of an E. coli wt and a mutant inactivated for RNase III
design: RNA-seq analysis of primary, processed and total RNAs of an E. coli wt and a mutant inactivated for RNase III
library_construction_protocol: Sampling of 10mL of bacteria grown to an A650=0.35–0.4 in LB medium at 37°C. Total RNA was ligated to the PSS RNA primer (Processing Start Site specific primer) After degradation of remaining 5'monophosphate RNAs. Treatment of the remaining RNAs by 5'polyphosphatase was followed by ligation of the TSS RNA primer (Transcription start specfic primer). RNAs were fragmented using the NEBnext Magnesium RNA fragmentation module and purified on an RNEasy column (Qiagen). Total RNA was extracted by the hot-phenol method then treated with DNase Library was built using the Truseq small RNA sample prep from Illumina.