RNA-Seq of Agave amacroacantha: leaf
The purity, concentration, nucleic acid absorbance peaks, and integrity of RNA are as-sessed using (Termo Fisher Scientifc, Waltham, MA, USA) and Agilent Bioanalyzer 2100 system (Agilent Technologies,Santa Clara, CA, USA), respectively. After obtaining high-quality RNA, library construction is carried out. First, mRNA is enriched, then mRNA is randomly fragmented using a fragmentation buffer; the first strand of cDNA is synthesized with random hexamers, followed by the synthesis of the second strand of cDNA in a conventional manner; the synthesized cDNA needs to be purified with AM-Pure XP beads, end-repaired, and sequencing adapters ligated; after that, AMPure XP beads are used again for size selection of the fragments; finally, the cDNA library is ob-tained through PCR.