title: RNAseq of 14-days cauliflower
design: Total RNA was extracted using the PureLink Plant RNA Reagent (Thermo Fisher Scientific) following the manufacturer's instructions. The extracted RNA was further purified using the Direct-zol RNA Miniprep Kit (Zymo Research) to enhance purity and integrity. RNA quality and concentration were assessed using a NanoDrop spectrophotometer (Thermo Fisher Scientific), the Qubit RNA Broad Range Assay Kit (Invitrogen), and an Agilent 2100 Bioanalyzer with the RNA 6000 Nano Kit (Agilent Technologies). For transcriptome sequencing, cDNA libraries were prepared using the NEXTflex Rapid Directional RNA-Seq Kit (Bioo Scientific) according to the manufacturer's protocol. Library quality was evaluated before sequencing. Sequencing was performed using the MiSeq Reagent Kit v3 on an Illumina MiSeq platform at WELGENE Biotechnology Company (Taipei, Taiwan).