title: GSM3414852: eds4 nuclear rep 1; Arabidopsis thaliana; RNA-Seq
library_construction_protocol: In the transcriptional profiling assay total RNA was extracted with RNeasy Plant Mini Kit (Qiagen) following the manufacturer's protocols. For the distribution of transcripts assay RNA was phenol-extracted, precipitated in sodium acetate, ethanol, and glycogen, washed in 80% ethanol and re-suspended in RNase-free water. Libraries were prepared following the TruSeq RNA Sample Preparation Guide (Illumina). Briefly, 3 μg of total RNA was polyA-purified and fragmented, first-strand cDNA synthesized by reverse transcriptase (SuperScript III; Invitrogen) using random hexamers. This was followed by RNA degradation and second-strand cDNA synthesis. End repair process and addition of a single A nucleotide to the 3′ends allowed ligation of multiple indexing adapters. Then, an enrichment step of 12 cycles of PCR was performed. Library validation included size and purity assessment with the Agilent 2100 Bioanalyzer and the Agilent DNA1000 kit (Agilent Technologies). Samples were pooled to create 12 multiplexed DNA libraries, which were pair-end sequenced with an Illumina HiSeq 1500.
attributes.tag: GEO Accession
attributes.value: GSM3414852