RNA Seq of Panicum virgatum: Genotype 421 lower half of 2nd internode rep 1
Greenhouse grown switchgrass samples were ground with liquid nitrogen by mortar and pestle. Each sample type has three biological replicates. Fine powderswere taken out for total RNA extraction by using RNeasy Plant Mini Kit (Qiagen) according to manufacturers instructions. All RNA quality was verified by  Agilent 2100 Bio-analyzer. The cDNA libraries were constructed by using TruSeq RNA library prep kit (Illumina). In brief, RNA samples were  treated with DNase I to remove DNA in RNA samples. Then, messenger RNA was isolated using poly (T) oligo-attached magnetic beads and fragmentized by covaris prior to the first cDNA strand synthesis with random hexamer primers. The second cDNA strand was synthesized to create a double-stranded cDNA for end-repair, 3 adenylation, adaptor ligation, and gel purification. Finally, cDNA libraries were enriched by polymerase chain reactions and qualified by Agilent Technologies 2100 Bio-analyzer and ABI StepOnePlus Real-Time PCR System. Samples were randomly distributed to flow cell lanes.
421_SL-1