title: RNA-Seq of  flower petals of Bombax ceiba
design: RNA was isolated utilizing a TRIzol reagent from Invitrogen, adhering to the specified protocols provided by the manufacturer. The RNA's integrity was assessed via the RNA Nano 6000 Assay Kit on the Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA). Following quantification, eukaryotic mRNA was selectively enriched using capture beads. Subsequently, single-stranded cDNA was synthesized employing six-base random primers, which was then converted into double-stranded cDNA. The resultant double-stranded cDNA was purified using the AMPure XP system (Beckman Coulter, Beverly, USA). The purified cDNA underwent end repair, adapter ligation, and size selection via DNA Clean Beads. After the adenylation of the 3' termini of the DNA fragments, adapters featuring a hairpin loop structure were incorporated to enhance hybridization efficiency. To isolate cDNA fragments within the range of 370 to 420 bp, a second purification step was performed using the AMPure XP system (Beckman Coulter, Beverly, USA). Following this, PCR amplification was conducted utilizing Phusion High-Fidelity DNA polymerase, in conjunction with universal PCR primers and the Index (X) Primer. The final PCR products were purified using the AMPure XP system, and the quality of the sequencing library was assessed using the Agilent Bioanalyzer 2100 system. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia) according to the manufacturer's instructions. After cluster generation, the library preparations were sequenced on an Illumina Novaseq platform and 150 bp paired-end reads were generated.