characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2019-03-17T17:02:52Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2016-03-17T12:06:58Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA3903308
characteristics.characteristic: External Id
characteristics.text: Cell 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: 2016-03-17T12:06:58Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1090442
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-4562:Fusarium_Medium_2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: Fusarium conidia were grown and cultivated in Erlenmeyer flasks that contain liquid complete medium as 25-40% of the total volume of the flask for 3 days on a rotary shaker at 175 rpm, at 28°C. The mycelium was then separated from the medium by filtration, washed up 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: media
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0000579
characteristics.characteristic: growth condition
characteristics.text: Fusarium graminearum
characteristics.characteristic: organism
characteristics.text: Fusarium graminearum
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_5518
characteristics.characteristic: organism
characteristics.text: Fusarium graminearum
characteristics.characteristic: scientific_name
characteristics.text: Fusarium_Medium_2
characteristics.characteristic: title