GSM4982272: paht mutant phe repl 3; Novosphingobium sp. HR1a; RNA-Seq
Total RNA was extracted using the Trizol method (TRIzol RNA Isolation Reagents, Thermofisher Scientist) and further DNAse treatment and purification with RNeasy Mini Kit (Qiagen). RNA degradation and contamination was monitored on 1% agarose gels. RNA purity was checked using the NanoPhotometer® spectrophotometer (IMPLEN, CA, USA). RNA integrity and quantification were assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). One μg RNA per sample was used to prepare the sequencing libraries which were generated by Novogen (Hong Kong) using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's recommendations and index codes were added to attribute sequences to each sample. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using PE Cluster Kit cBot-HS (Illumina) according to the manufacturer's instructions. After cluster generation, the library preparations were sequenced on an Illumina platform and paired-end reads were generated.