Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNA was extracted using the RNEasy Plant Mini Kit (Qiagen, Carlsbad, CA) and treated with Turbo DNase (Life Technologies, Carlsbad, CA) to remove contaminating genomic DNA. Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit (Part# 0801-0303). Libraries were prepared using the NEBNext mRNA Library Prep Master Mix Set for Illumina and according to kit's instructions. Briefly, polyA mRNA was purified from 1 µg total RNA using the Dynabeads mRNA DIRECT Purification kit. After mRNA fragmentation and cDNA synthesis, adaptors were ligated and library fragments of ~100 bp were selected using AMPure XP beads. DNA fragments were PCR amplified with Illumina primers for 10 cycles. Libraries were sequenced on the Illumina NextSeq 500 platform following the manufacturer's protocols. RNA-seq each library was split into four parts for sequencing on independent lanes so as to remove any lane bias