SoxR-G121P-3
Total RNA of each sample was extracted by RNeasy Mini Kit (Qiagen, Valencia, CA, USA) after the cells of positive variant strains were grown to mid-exponential phase at 37 C and 220 rpm. The quantity and quality of RNA were verified using a Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA). Total RNA (1 g) with an RNA Integrity Number (RIN) value above 6.5 was used for following library preparation. Next generation sequencing library preparations were constructed according to the manufacturers protocol (NEBNext Ultra RNA Library Prep Kit for Illumina). The rRNA was depleted from total RNA using rRNA removal Kit. The ribosomal depleted RNA was then fragmented and reverse-transcribed to synthesis cDNA. Then libraries with different indices were multiplexed and loaded on an Illumina HiSeq instrument according to manufacturers instructions (Illumina, San Diego, CA, USA).