GSM5057838: ptsI 90 min; Escherichia coli K-12; RNA-Seq
Total RNAs were prepared using an RNA extraction kit following the manufacturer protocol (Tiangen, China). A total amount of 3 µg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB, USA) following manufacturer's recommendations. Briefly, rRNAs were removed by the Ribo-zero kit to enrich mRNAs, which were then digested into short fragments. The fragmented mRNAs were used as template to generate one-strand DNA using a six-base random primer and the corresponding double-stranded cDNA. After purification, the cDNA was firstly end-repaired, A-tailed and ligated to a sequencing linker. The size of fragments was selected using AMPure XP beads. PCR amplification was performed, and the amplified products were then purified using AMPure XP beads to obtain the final library. The cDNA library was sequenced (RNA-seq) on an Illumina novaseq platform, and paired-end reads were generated.