RNA-Seq of Juglans sigillata Dode female flower bud Undifferentiated stage Experiment Replicate 1
Qualified total RNA was first purified and fragmented randomly using divalent cations under elevated temperature. The fragmented RNA was then reverse transcribed into cDNA using random primers. The cDNA was end-repaired, A-tailed, and adaptor ligated using the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, USA) according to the preparation protocol. The ligated products were subjected to size selection to enrich fragments of 300-400 bp. Enrichment of cDNA fragments was carried out by PCR amplification. Finally, PCR products were purified using the AMPure XP system (Beckman Coulter, Brea, CA, USA). The libraries were analyzed for size distribution using the 2100 Bioanalyzer (Agilent, Santa Clara, CA) and quantified using real-time PCR. RNA sequencing was performed on the Illumina NovaSeq X Plus sequencer using paired-end technology (PE 150).