RNA-seq of roots of Arabidopsis under full P condition
Total RNA from the samples was extracted using FavorPrepTM plant total RNA minikit (Favorgen), and cDNA libraries corresponding to the treated and control (untreated) plants were constructed using TruSeq Stranded mRNA LT Sample Prep Kit (Illumina, USA). Briefly, poly-A containing mRNA molecules were purified using poly-T oligo-attached magnetic beads. Following purification, the mRNA was fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNA fragments were then used to synthesize first strand cDNA using reverse transcriptase and random primers followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. These cDNA fragments were then subjected to an end repair process, followed by the addition of a single A base, and then ligation of the adapters. The products were purified and enriched with PCR to create the final cDNA library. Bioanalyzer plots were used to assess mRNA quality, enrichment success, fragmentation sizes, and final library sizes.