RNA-Seq of Watermelon line: Leaf
Total RNA was isolated from frozen true leaf tissues using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) following manufacturers instructions. RNA purification steps and on-column DNase digestion was performed using the QIAGEN RNeasy Mini Kit as suggested by the manufacturers instructions (QIAGEN, Hilden, Germany). The total RNA qualities were further analyzed using Agilent RNA 6000 Nano Kit (Agilent Technologies Inc., Santa Clara, CA, USA) ran on a Agilent 2100 Bioanalyzer, with average RNA RIN values>7. RNA-seq libraries were prepared as described previously25. The quality and integrity of each library was checked with Agilent high-sensitivity DNA chips (Agilent Technologies, Santa Clara, CA, USA). Concentrations of sequencing libraries were quantified using Qubit fluorometer and Qubit dsDNA HS assay kit (Life Technologies, CA, USA). Three biological replications of each genotype (USVL531-MDR, USVL677-PMS) per time-point were used for 150 bp paired-end RNA-seq sequencing (Duke Center for Genomic and Computational Biology, Durham, NC, USA) with Illumina HiSeq 4000 sequencing system (Illumina, Inc., San Diego, CA, USA).