GSM6696303: Cu_2; Cupriavidus basilensis; RNA-Seq
RNA was extracted using a phenol-chlorform method. Briefly, 500 µL culture and 1 ml PBS are centrifuged at 5000 g for 5 min. Lysozyme and Superase-IN RNase inhibitor are used. This is followed by bead-beating with RNASolv reagent (Omega Bio-tek). Chloroform is added and the samples sit at room temperature for 3 min. The samples are then centrifuged at >12000 g for 15 min at 4 ºC. The top aqueous layer is removed and an equal volume of 70% ethanol is added. The samples are then processed using the EZNA Total RNA Kit according to the manufacturer's instructions (Omega Bio-Tek). 5 µl of each sample (concentration 2-10 ng/µl) was used for library preparation with the NEBNext Ultra II Directional RNA library Prep kit following the manufacturer's instructions. NovaSeq platform for paired-end 150bp 