RNA-seq of Eucommia ulmoides: leaf under normal temperatures
To extract total RNA from E. ulmoides, the Trizol method was employed. The purity of the RNA was assessed using the NanoDrop method, calculating the OD260/OD280 ratio, and its quality was verified using 1% agarose gel electrophoresis. Subsequently, Oligo (dT) beads were utilized to isolate the mRNA, followed by fragmentation of the mRNA into short fragments using an interrupting reagent. These short fragments were employed as templates for the synthesis of the first cDNA strand via first-strand synthesis. Following this, the Second Strand Synthesis Mix was utilized to synthesize the second strand. The resulting product underwent end leveling and had A added to it. RNA-Seq Adapter was connected to the product after this process. After ligating the RNA-Seq Adapter, the ligated product was purified, followed by PCR amplification. The target product of 300600 bp was purified and recovered using magnetic beads. Finally, the cDNA library was sequenced on the Illumina NovaSeq 6000 platform once the library construction was deemed qualified