Torreya grandis leaf transcriptome: male, 6d PEG treatment, rep1
Total RNA was evaluated for quality. Poly(A) mRNA was enriched using oligo(dT) magnetic beads, fragmented, and reverse-transcribed into first-strand cDNA using random hexamers. Second-strand cDNA was synthesized using RNase H and DNA Polymerase I. The double-stranded cDNA was purified using a QIAquick PCR kit, end-repaired, A-tailed, and ligated to sequencing adapters. After PCR amplification, products were denatured and circularized to generate single-stranded circular DNA. DNA nanoballs (DNBs) were prepared and sequenced on the MGI platform with a PE150 strategy.