GSM9316567: ΔrecA_MMC_0_replicate1; Caulobacter vibrioides; RNA-Seq
The samples were then centrifuged at 10000g for 5 minutes following which supernatant was discarded and the pellets were snap frozen and stored at -80°C until further processed. mRNA was isolated from the collected pellets. Pre-heated trizol (65°C) was added to each pellet and cells were lysed using a Thermomixer (65°C, 2000rpm, for 10 minutes). Total RNA was extracted from the lysed cells using the Direct-zol RNA MiniPrep kit. Any DNA contamination was then removed via DNAse treatment and  RNA was purified using RNA Clean & Concentrator-25 kit. Integrity of the total RNA was tested by the Tapestation instrument. Ribosomal RNA was then depleted using the Thermo RiboMinus kit. mRNA was purified using RNA Clean & Concentrator-5 kit and submitted for RNA-seq. NEBNext Ultra II Directional RNA Library Prep kit was used for library preparation and samples were sequenced via the Illumina NovaSeq 6000 platform.