GSM9035180: L. lactis subps. cremoris MG1363_sLLM2-_rep2; Lactococcus cremoris subsp. cremoris MG1363; RNA-Seq
Bacteria were mixed with 1/8 volume of ice-cold STOP solution (95% ethanol and 5% acid phenol) and centrifuged at 1,800 × g for 20 min at 4°C. Cells were ground to a fine powder using liquid nitrogen and total RNA was extracted with RNAzolRT (RN 190) (Molecular Research Center, Inc.). Total RNA was treated with DNase I, RNase-free (Thermo Scientific), and purified using RNA Clean & Concentrator-5 Kit (Zymo Research). rRNA was depleted using the modified RiboMinus Transcriptome Isolation Kit (Invitrogen), bacteria, where 3,5 μl of commercial probes and 0,5 μl of a 100 μM mix of three custom-designed probes specific to L. lactis 5S rRNA were used. After ethanol precipitation, libraries were further prepared in ThermoFisher Scientific facilities using Invitrogen™ Collibri™ Stranded RNA Library Prep Kit for Illumina.