RNA-seq of Vibrio alginolyticus: Del_hfq-T
"The bacterial cells were grown in TSB medium at 30deg C to early log phase (OD600 is 0.5). Total RNA was isolated using RNAiso Plus (Takara) and treated with DNase I, then removed rRNAs. Mixed with the fragmentation buffer, the mRNA was fragmented into short fragments. Then cDNA was synthesized using the mRNA fragments as templates. Short fragments were purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments were connected with adapters. After agarose gel electrophoresis, the suitable fragments were selected for the PCR amplification as templates. During the QC steps, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System were used in quantification and qualification of the sample library. At last, the library was sequenced using Illumina HiSeq 2000."