GSM3169153: root_96hours_control_rep2; Lens culinaris; RNA-Seq
Total RNA was isolated from 1 gr of root, stem and leaves of Lens culinaris using TRIzol reagent (Invitrogen, USA) according to the manufacturer's instructions. BioAnalyzer (Agilent, USA) was used to measure the quality and the quantity of the isolated total RNAs. After that, total RNA samples were treated with RNase-free DNase I (Thermo Scientific, USA) at a concentration of 1 U/µg to eliminate the genomic DNA. Then, RNA-seq library preparation was performed using TruSeq Stranded RNA LT Kit (Illumina, USA) according to the manufacturer's instructions. mRNAs were purified from the 1 µg of total RNA using oligo (dT) magnetic beads and fragmented using fragmentation buffer. Afterthat, the cleaved short RNA fragments were used for first-strand cDNA synthesis using first strand synthesis mix, and the second strand was synthesized using second strand marking master mix. The double strand cDNAs were purified with AMPure XP beads (Beckman Coulter, USA) and eluted with resuspension buffer followed by 3'end adenine nucleotide addition. Finally, sequencing adaptors were ligated to the fragments and cDNA fragments were enriched by PCR amplification. Enriched cDNA libraries were used for cluster generation and sequencing. 100x2 paired-end sequencing of cDNA libraries for control and drought stress exposed samples (1st day and 4th day samples of root and leaf, and 1st day sample of stem) with two biological replicates were performed using the Illumina HiSeq4000 sequencing platform (Illumina, USA).
GEO Accession
GSM3169153