characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2019-03-17T17:02:52Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2016-03-15T16:45:48Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA3899920
characteristics.characteristic: External Id
characteristics.text: GermanyCell Biology & Plant Biochemistry University of Regensburg
characteristics.characteristic: INSDC center name
characteristics.text: 2019-03-17T17:02:52Z
characteristics.characteristic: INSDC first public
characteristics.text: 2016-03-15T16:45:48Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1087054
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-4559:Leaf_2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: maize
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PO_0020136
characteristics.characteristic: common name
characteristics.text: Protocols: Plants were grown in a greenhouse under controlled conditions with 16 hours light of 26-28°C and 8 hours night of 18-20°C. Third leaves of 21 days old plants were harvested and stored at -80°C. Total RNA was prepared by grinding tissues in liquid nitrogen and total RNA was extracted by TRIzol reagent (TRIzol Plus RNA Purification Kit, Ambion) and processed following the protocol provided by manufacturer. To remove DNA within extraction and purification procedures, total RNA was treated with DNase (Qiagen), then total RNA was quantified and visualized on a Bioanalyzer (Agilent). Library preparation and RNAseq were carried out as described in the Illumina TruSeq RNA Sample Preparation v2 Guide, the Illumina HiSeq 1000 System User Guide (Illumina, Inc., San Diego, CA, USA), and the KAPA Library Quantification Kit - Illumina/ABI Prism User Guide (Kapa Biosystems, Inc., Woburn, MA, USA). In brief, 500 ng of total RNA was used for purifying the poly-A containing mRNA molecules using poly-dT oligo-attached magnetic beads. Following purification, the mRNA was fragmented to an average insert size of 200-400 bases using divalent cations under elevated temperature (94°C for 4 minutes). The cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random primers, followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. The resulting cDNA fragments subsequently went through an end repair process, the addition of a single ‘A’ base, the ligation of the adapters, and a purification step. Finally the cDNA libraries were created by PCR enrichment. The libraries were quantified using the KAPA SYBR FAST ABI Prism Library Quantification Kit. Equimolar amounts of each library were used for cluster generation on the cBot (TruSeq PE Cluster Kit v3).
characteristics.characteristic: description
characteristics.text: Zea Mays_Inbred B73
characteristics.characteristic: genotype
characteristics.text: Zea mays subsp. mays
characteristics.characteristic: organism
characteristics.text: Zea mays subsp. mays
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_381124
characteristics.characteristic: organism
characteristics.text: leaf
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PO_0025034
characteristics.characteristic: organism part
characteristics.text: Zea mays subsp. mays
characteristics.characteristic: scientific_name
characteristics.text: Leaf_2
characteristics.characteristic: title