title: "Miscanthus lutarioriparius transcriptome project,the mature leaf sample of LU_DS_14-5"
design: Total RNA of leaves was extracted using Qiagen Plant Mini Kit (Qiagen, Stanford, CA, USA) and purified using the RNase-free DNaseⅠ(TaKaRa) following the manufacturer’s protocol.  The concentration of purified RNA was quantified by NanoDrop ND-1000 spectrophotometer (Thermo Scientific, Wilmington, DE, USA) and stored at -80℃ until next step.  The mRNA was isolated from 5μg purified total RNA using one round of purification with oligo d(T) beads Dynabeads? mRNA Purification Kit (Invitrogen).  The cDNA libraries were prepared with the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs, MA, USA).  The first strand cDNA was synthesized using random hexamer-primed reverse transcription.  Following the second strand cDNA synthesis and adaptor ligation, approximately 450-bp cDNA fragments were isolated by the selection of Ampure XP beads (Beckman).  The isolated cDNA fragments were amplified by 10 cycles of PCR.  Library integrity and quality were estimated with Agilent 2100 Bioanalyzer (Agilent Technologies) and Qubit 2.0 fluorometer (Life Technologies) respectively.  The 100-bp paired-end sequencing was sequenced on an Illumina HiSeq 2500.