Lmin2
After total RNA was extracted, eukaryotic mRNA wasenriched by Oligo(dT) beads, while prokaryotic mRNA was enriched by removingrRNA by Ribo-ZeroTM Magnetic Kit (Epicentre, Madison, WI, USA). Then the enrichedmRNA was fragmented into short fragments using fragmentation buffer and reversetranscribed into cDNA with random primers. Second-strand cDNA was synthesized byDNA polymerase I, RNase H, dNTP and buffer. Then the cDNA fragments werepurified with QiaQuick PCR extraction kit(Qiagen, Venlo, The Netherlands), endrepaired, A base added, and ligated to Illumina sequencing adapters. The ligationproducts were size selected by agarose gel electrophoresis, PCR amplified, andsequenced using Illumina novaseq 6000 by Gene Denovo Biotechnology Co.(Guangzhou, China)