GSM5219475: 10CEB13BAC -1 [GLA-5_S8]; Bacillus cereus; RNA-Seq
The bacterial pellets were re-suspended with 200 μl of 10 mM Tris-HCl at pH 8 + 4 μl of lysozyme at 50 mg/ml and incubated at 37°C. Total RNA was extracted with the HPRNA kit (High Pure RNA Isolation Kit; Roche). Remaining traces of DNA were removed by two steps of DNase treatment (TURBO-DNAse Free kit, Ambion). The RNA integrity was measured by the RIN (RNA Integrity Number) obtained using the capillary electrophoresis Bioanalyser® (Agilent). RIN was calculated with the total RNA ratio of the area of ribosomal bands to the total area and the height of the 16S and 23S peaks. The RIN of samples were between 7 and 10. The mRNA were purified with the RiboZero Kit (Illumina). Directional and paired libraries were prepared with the Illumina scriptseq kit and the sequencing was performed on an Illumina Nextseq machine.