title: RNA-Seq of GmALA1 overexpressed transgenic roots
design: After enriching eukaryotic mRNA with polyA tails using magnetic beads with Oligo(dT), the mRNA is fragmented using ultrasound. Taking the fragmented mRNA as a template and random oligonucleotides as primers, the first strand of cDNA is synthesized in the M-MuLV reverse transcriptase system. Subsequently, the RNA strand is degraded using RNase H, and the second strand of cDNA is synthesized using DNA polymerase I with dNTPs as substrates. The purified double-stranded cDNA undergoes end repair, A-tailing, and ligation of sequencing adapters. AMPure XP beads are used to select cDNA fragments around 200bp, which are then amplified by PCR. The PCR products are purified again using AMPure XP beads to obtain the final library.