title: GSM7819312: dek2_Treated_H10; Arabidopsis thaliana; RNA-Seq
library_construction_protocol: 50-100 mg of Arabidopsis plant leaves or 14 days old seedling grown on ½ MS plates were taken in 2 ml tubes containing 2 steel beads, frozen in liquid Nitrogen and homogenized 2X using a Tissue Lyser (Eppendorf) for 1 min at 20 Hz to obtain a fine powder.  NucleoSpin® Plant RNA kit (Macherey Nagel, 7740949) was used to extract the total RNA from the plant. mRNA libraries were prepared using the illumina Truseq Stranded mRNA Sample Preparation LS (low sample) kit following the manufacturer's protocol. Briefly, the mRNA was purified from 1ug of total RNA using poly-T oligo-attached magnetic beads then purified, fragmented and primed. Subsequently, both first and second strands of cDNA were synthesized using SuperScript II reverse transcriptase, followed by adenylation on the 3' ends and ligation of adapters. The DNA fragments with adapters were amplified by 15 PCR cycles. Finally, the libraries were validated using the 1000 DNA kit on 2100 Bioanalyzer (Agilent Technologies), quantified using qubit (Life Technologies) then barcoded libraries were normalized and pooled in equal volumes. Stranded mRNA libraries were sequenced on an Illumina Hiseq 4000 system using paired-end method. The length of the read was around 150 bp. The RNA-seq was done on three independent biological replicates.