title: RNA-seq of Cannabis sativa under drought stress
design: The acquisition of mRNA can be achieved by two principal approaches. The first exploits the characteristic polyadenylated tail present on the majority of eukaryotic mRNAs: poly(A)-tailed transcripts are selectively enriched using Oligo(dT) magnetic beads. The second entails depletion of ribosomal RNA from total RNA preparations, thereby yielding an mRNA-enriched fraction. Subsequently, fragmentation buffer is added to cleave the RNA into short fragments. These fragmented RNAs serve as templates for first-strand cDNA synthesis, which is primed by random hexamers. Following this, buffer, a complete set of deoxynucleotide triphosphates (dTTP, dATP, dGTP, and dCTP), and DNA polymerase I are introduced to synthesize the second cDNA strand. The resulting double-stranded cDNA is then purified using AMPure XP beads. The purified double-stranded cDNA undergoes end repair, A-tailing, and ligation of sequencing adapters. Adapter-ligated fragments are size-selected with AMPure XP beads, and the final cDNA library is obtained by PCR amplification.