Illumina HiSeq 2500 paired end sequencing; RNA-Seq of Clostridium cellulovorans cultured in different carbon sources in exponential phase
RNA-Seq of Clostridium cellulovorans cultured in different carbon sources in exponential phase
Cell pellets from 50 ml culture were collected by centrifugation at -4 °C and 9,000 rpm (Hitachi CF16RXII, Qingdao, China) for 5 min. Afterwards, pellets were frozen in liquid nitrogen and stored at -80°C. Total RNA was extracted using the mirVana miRNA Isolation Kit (Ambion) following the manufacturer's protocol. RNA integrity was evaluated using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The samples with RNA Integrity Number (RIN) ≥ 7 were subjected to the subsequent analysis. Double stranded cDNA was generated using the SuperScript Double-Stranded cDNA synthesis kit (Invitrogen) according to the manufacturer's instructions.It was performed by Oebiotech (Shanghai, China). The libraries were constructed using TruSeq Stranded mRNA LTSample Prep Kit (Illumina, San Diego, CA, USA) according to the manufacturer's instructions. This part of experiment is carried out at Oebiotech (Shanghai, China).