RNA-seq of EV_1 transgenic roots
Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. After library quality control, different libraries were pooled based on the effective concentration and targeted data amount. 5'-end of each library was phosphorylated and cyclized. Subsequently, loop amplification was performed to generate DNA nanoballs. These DNA nanoballs were finally loaded into flowcell with DNBSEQ-T7 for sequencing.