GSM9194767: B46,Maize,rep1; Zea mays; RNA-Seq
Total RNA was extracted following the standard TRIzol protocol (Invitrogen, Carlsbad, CA) as described by Lemmon (Lemmon et al., 2014, 10.1371/journal.pgen.1004745). Transcriptome library preparation and sequencing were performed by Novogene Co., Ltd. (Beijing, China). Briefly, polyadenylated mRNAs were enriched using Oligo(dT) magnetic beads and randomly fragmented in NEB Fragmentation Buffer containing divalent cations (Parkhomchuk et al., 2009). First-strand cDNA synthesis was carried out using random oligonucleotide primers and M-MuLV reverse transcriptase, followed by second-strand synthesis using DNA polymerase I and dNTPs after RNaseH digestion. Purified double-stranded cDNA underwent end-repair, A-tailing, and adapter ligation. Fragments of approximately 370–420 bp were selected using AMPure XP beads, PCR-amplified, and further purified to finalize the libraries.