title: GSM1930763: Col-0 WT Rep1; Arabidopsis thaliana; RNA-Seq
library_construction_protocol: RNA from inflorescences were extracted with the RNeasy mini kit (Qiagen) followed by Dnase treatment. Sequencing was done at the Zurich Functional Genomics Center (www.fgcz.ch). The quality of the isolated RNA was determined with a Qubit® (1.0) Fluorometer (Life Technologies, California, USA) and a Bioanalyzer 2100 (Agilent, Waldbronn, Germany). Only those samples with a 260 nm/280 nm ratio between 1.8–2.1 and a 28S/18S ratio within 1.5–2 were further processed. The TruSeq Total RNA Sample Prep Kit v2 (Illumina, Inc, California, USA) was used in the succeeding steps. Briefly, total RNA samples (100-1000 ng) were ribo-depleted using Ribo Zero (Epicentre®, USA)  and then reverse-transcribed into double-stranded cDNA. The cDNA samples was fragmented, end-repaired and polyadenylated before ligation of TruSeq adapters containing the index for multiplexing Fragments containing TruSeq adapters on both ends were selectively enriched with  PCR.  The quality and quantity of the enriched libraries were validated using Qubit® (1.0) Fluorometer and the Caliper GX LabChip® GX (Caliper Life Sciences, Inc., USA).  The product is a smear with an average fragment size of approximately 260 bp. The libraries were normalized to 10nM in Tris-Cl 10 mM, pH8.5 with 0.1% Tween 20. The TruSeq PE Cluster Kit v3-cBot-HS (Illumina, Inc, California, USA) was used for cluster generation using 10 pM of pooled normalized libraries on the cBOT.  Sequencing were performed on the Illumina HiSeq 2000 paired end at 2 X101 bp using the TruSeq SBS Kit v3-HS (Illumina, Inc, California, USA).
attributes.tag: GEO Accession
attributes.value: GSM1930763