title: Illumina HiSeq 2000 sequencing; Multifaceted Activity of Cytokinin at Distinct Phases of Leaf Development Shapes Its Size and Structure in Arabidopsis thaliana
design: Multifaceted Activity of Cytokinin at Distinct Phases of Leaf Development Shapes Its Size and Structure in Arabidopsis thaliana
library_construction_protocol: Collection of plant materials from in-vitro growing Arabidopsis plants.  The leaf 3 was dissected from i > 250 individual seedlings per variant on a cooling plate under a stereomicroscope with precision needle and removed into the liquid nitrogen. DEX-inducible lines and wild-types were activated at 9 DAS by transfer of nylon meshes to half-strength MS medium supplemented with either 2.5 M DEX dissolved in 5x10-4% (v/v) DMSO or just 5x10-4% (v/v) DMSO. Poly(A)-containing mRNA molecules were reverse transcribed, double-stranded cDNA was generated and adapters were ligated. The total RNA was extracted according to a combined protocol of TRI Reagent RT (Molecular Research Center) and the RNeasy Kit (Qiagen) with on-column DNase (Qiagen) digestion. The TruSeq RNA Sample Preparation Kit version 2 (Illumina) was used for library preparation.
attributes.tag: Experimental Factor: phenotype
attributes.value: dexamethasone-inducible expression of barley cytokinin oxidase/dehydrogenase