title: GSM3693408: ctr1_3; Vitis vinifera; RNA-Seq
library_construction_protocol: Stem tissue samples were ground in liquid nitrogen and total RNA was extracted using CTAB-based extraction protocol according to Gambino et al. (2008) followed by a DNase I treatment (RNase-Free DNase Set, Qiagen). Concentration and quality of the total extracted RNA were checked using the 'Quant-it ribogreen RNA assay' (Life Technologies, Grand Island, NY, USA) and the RNA 6000 nano chip (Agilent Technologies, Santa Clara, CA, USA), respectively. Illumina mRNA sequencing libraries were made from 200 ng of total RNA of each sample using the QuantSeq 3' mRNA-Seq Library Prep Kit (Lexogen, Vienna, Austria) according to manufacturer's protocol using 15 cycles for the enrichment PCR step. Libraries were quantified by qPCR on a Lightcycler 480 (Roche, Basel, Switzerland), according to Illumina's protocol 'Sequencing Library qPCR Quantification protocol guide', version February 2011. A High Sensitivity DNA chip (Agilent Technologies, Santa Clara, CA, US) was used to control the library's size distribution and quality. The libraries were equimolarly pooled and sequenced at the NXTGNT massively parallel sequencing facility (www.nxtgnt.ugent.be, Ghent University) on an Illumina NextSeq 500 high throughput flow cell, generating single-end 75bp reads.
attributes.tag: GEO Accession
attributes.value: GSM3693408