GSM2701803: Dh2; Citrus sinensis; RNA-Seq
Total RNA was extracted from frozen AZ-C by RNeasy Plant Kit (Qiagen) following the manufacturer’s instructions. DNA was removed by on column DNase treatment with RNase-free DNase Set (Qiagen). The purity and concentration of total RNA were determined using a 2100 bioanalyzer (Agilent). cDNA libraries were generated using the mRNA Sequencing Sample Preparation Kit following the manufacturer’s instructions (Illumina, CA, USA). Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads and cDNA was then synthesized. cDNA fragments 200 ± 25 bp in size were selected and PCR was performed with High-Fidelity DNA polymerase, universal PCR primers and Index (X) Primer. PCR products were purified and the library preparations were sequenced on the Illumina cluster station and Illumina HiSeq1500 sequencing platform.