"Miscanthus lutarioriparius transcriptome project,the mature leaf sample of LU7-20"
"We sampled 80 individuals (8 individuals per population) from ten populations. Total RNA of each individual was isolated using Qiagen Plant Mini Kit (Qiagen), purified using the RNase-free DNase I (TaKaRa), quantified by Nanodrop 1000 instrument (Thermo Scientific) and stored at -80 °C. The mRNA of each individual was isolated from 5μg of 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). 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. The completeness and quantification of the library were assessed by Agilent 2100 Bioanalyzer (Agilent Technologies) and Qubit? 2.0 fluorometer (Life Technologies), respectively. The 100-bp paired-end sequencing was performed on an Illumina HiSeq 2500."
Institute of Botany, the Chinese Academy of Scienc
LU7-20