Leymus arenarius transcriptome, normal conditions
Total RNA was isolated from 5g of fresh 14-old-day seedlings using the RNAwiz kit (Ambion, Canada) according to the manufacturer’s instructions. Half of the total RNA sample was used for cDNA library preparation for Illumina Sequencing, and half of the sample -  for RT-PCR validation.   Library Preparation and Illumina Sequencing  The quality of total RNA was checked using a NanoDrop 2000 (Thermo Fisher, USA) and an Agilent 2100 Bioanalyzer (Agilent, USA). mRNA was purified from total RNA by RNA purification beads, then fragmented and primed for cDNA synthesis according to the manufacturer’s instructions (Illumina, USA). Double-stranded cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, USA) and then purified using Agencourt AMPure XP beads (Beckman Coulter, Inc, USA). End repairing and 3’-ends adenylation were performed following the RNA adapters ligation. After enrichment of DNA fragments library templates were validated using the Aligent 2100 Bioanalyzer (Agilent, USA). Using TruSeq PE Cluster Kit v2 and cBot automated system (Illumina, USA) clonal clusters were created from DNA library templates. Obtained clusters were finally used to perform paired-end runs by Genome Analyzer IIx (Illumina, USA)
Vavilov Institute of General Genetics, Russian Aca
Leymus arenarius