GSM8014858: ATCC14579, Etest; Bacillus cereus; RNA-Seq
Total RNA was extracted using the RNeasy Protect Bacteria Mini Kit.  Approximately 1.8 OD600 of cells were used and all the RNA harvested was used to build the library. Total RNA was further purified using the DNA-free Kit (ThermoFisher). Bacterial ribosomal RNA was depleted using the NEBNext rRNA Depletion Kit Bacteria (NEB) followed by polyadenylating with E. coli poly(A) polymerase. The direct cDNA Kit (Oxford Nanopore) was used for reverse transcription and library construction. The Native Barcoding Expansion 1-12 (Oxford Nanopore) was used for multiplexing the cDNA samples. All steps were carried out according to the manufacturers protocol. Quantification and quality control of RNA and cDNA were reduced to a minimum to obtain sufficiently high amounts of RNA or cDNA.