GSM8121269: ONtreatedlD7_2; Allium cepa; RNA-Seq
Total RNA was extracted from each sample using the QIAseq fastSelect rRNA kits. The mRNA was isolated from the total RNA using magnetic beads with Oligo (dT) and fragmented into short segments using a fragmentation buffer. cDNA was synthesized via reverse transcription (Superscript III, Invitrogen). Oligo dT beads were used to enrich mRNA with poly A tail from total RNA, and mRNA molecules were fragmented into small pieces. The fragmented mRNA was used to first-strand cDNA using random primers, and the second-strand cDNA was synthesized with dUTP instead of dTTP. The synthesized cDNA was subjected to end-repair and 3' adenylation. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments. The U-labeled second-strand template was digested with Uracil-DNA-Glycosylase (UDG) before PCR amplification. The library was amplified to make DNA nanoball (DNB), and the sequencing was performed on the DNBSEQ (DNBSEQ Technology) platform.