GSM6927026: Escherichia coli C227-11 - 0 weeks after inoculation, rep3; Escherichia coli; RNA-Seq
RNAprotect Bacteria (Qiagen), following the manufacturer´s instruction. RNeasy mini kit (Qiagen), 700 µl of lysate with 470 µl of absolute ethanol. Followed by RNAase-free DNase set (Qiagen)+centrifugation steps to prevent DNA and ethanol contamination. A total of 1 µg of total RNA was subjected to rRNA depletion using Ribo-Zero rRNA Removal Kit Bacteria (Illumina). The cDNA libraries were constructed using the resulting tRNA and the NEBNext Ultra II Directional RNA Library Prep Kit (NEB). Libraries were sequenced as single-read (75 bp read length) on an Illumina NextSeq500 platform at a depth of 8–10 million reads