characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-09-29T17:01:53Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-08-10T14:12:56Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104209664
characteristics.characteristic: External Id
characteristics.text: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid (UPM) - Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA)
characteristics.characteristic: INSDC center name
characteristics.text: 2017-09-29T17:01:53Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-08-10T14:12:56Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1868682
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5963:WTR2
characteristics.characteristic: Submitter Id
characteristics.text: 7
characteristics.unit: day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: domesticated barley
characteristics.characteristic: common name
characteristics.text: Golden Promise
characteristics.characteristic: cultivar
characteristics.text: Protocols: Barley plants of Hordeum vulgare cv. Golden Promise were used. Grains were germinated in soil and grown at 22ºC under a 16 h light/8 h darkness photoperiod for 7 days in Sanyo MLR-350-H chambers. Barley transgenic lines overexpressing or silencing the HvPap-1 gene (OE Pap1 and KD Pap1, respectively) were generated using Agrobacterium-mediated gene transfer on immature embryos and homozygous material obtained by double haploid technology. Transgenic barley plants ubiquitously over-expressing the HvPap-1 gene were generated using p6U and p6d35s binary vectors. Silencing of the HvPap-1 gene was performed using the artificial microRNA (amiRNA) technology. Total RNA was extracted from frozen barley leaves by the phenol/chloroform method, followed by precipitation with 8 M LiCl and digested with DNase (Promega). Using poly-T oligo-attached magnetic beads, mRNAs were purified from the total RNA. Then, the mRNAs were fragmented and cDNA was synthesized using random hexamer-primers, DNA polymerase I and RNase H. The double-stranded cDNAs were purified with magnetic beads and ligated to adaptors for Illumina sequencing. The quality and quantity of the library was verified using an Agilent 2100 Bioanalyzer and an ABI StepOnePlus Real-Time PCR system, respectively.
characteristics.characteristic: description
characteristics.text: wild type genotype
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: genotype
characteristics.text: Hordeum vulgare subsp. vulgare
characteristics.characteristic: organism
characteristics.text: Hordeum vulgare subsp. vulgare
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_112509
characteristics.characteristic: organism
characteristics.text: leaf
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PO_0025034
characteristics.characteristic: organism part
characteristics.text: Hordeum vulgare subsp. vulgare
characteristics.characteristic: scientific_name
characteristics.text: WTR2
characteristics.characteristic: title