characteristics.text: 2012-10-26T17:01:43Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T15:47:11Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA1557047
characteristics.characteristic: External Id
characteristics.text: UCL-Gen
characteristics.characteristic: INSDC center name
characteristics.text: 2012-10-26T17:01:43Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T15:47:11Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS146820
characteristics.characteristic: SRA accession
characteristics.text: JB22
characteristics.characteristic: StrainOrLine
characteristics.text: E-MTAB-1154:MN1pA
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: fission yeast
characteristics.characteristic: common name
characteristics.text: Protocols: Cells were harvested from liquid cultures at mid-exponential phase (OD = 0.5). Cells were grown in Yeast Extract medium + Supplements (YES; Moreno et al., 1991, Methods Enzymol. 194, 795-823.) or in Edinburgh Minimal Medium (EMM; Moreno et al., 1991, Methods Enzymol. 194, 795-823.) at 32C. Removal of the nitrogen source from the medium (EMM minus nitrogen) the cells arrest in G1 and enter a resistant stage quiescence. Total RNA was isolated from S. pombe cells using a hot phenol method followed by phenol-chloroform extractions, and precipitation as described (Lyne et al., 2003, BMC Genomics 4, 27.). 10ug total RNA was used as starting material for strand-specific RNA-seq library preparation using an unreleased early version provided by Illumina of the Illumina TruSeq Small RNA Sample Prep Kit. Briefly: poly(A)+ RNA was enriched by two rounds of poly(dT) Sera-Mag magnetic beads purification. Enriched poly(A)+ RNA was then fragmented to an average size of 200nt. Fragmented RNA was 3prime de-phosphorylated with Antartic phosphatase and 5prime phosphorylated with polynucleotide kinase this treatment prepares RNA fragments for subsequent ligation of Illumina RNA adaptors to their 5prime and 3prime ends using a 3prime RNA ligase and a T4 RNA ligase, respectively. First strand cDNA was produced using a primer specific for the Illumina 3prime adaptor. The library was finally amplified with 15 cycles of PCR using primers specific for the Illumina adaptors and purified using SPRI-beads (Agencourt, Beckman Coulter). Libraries size distributions and concentrations were determined on an Agilent Bioanalyzer
characteristics.characteristic: description
characteristics.text: 972 h-
characteristics.characteristic: genotype
characteristics.text: Schizosaccharomyces pombe
characteristics.characteristic: organism
characteristics.text: Schizosaccharomyces pombe
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_4896
characteristics.characteristic: organism
characteristics.text: Schizosaccharomyces pombe
characteristics.characteristic: scientific_name
characteristics.text: mating_type_h_minus
characteristics.characteristic: sex
characteristics.text: MN1pA
characteristics.characteristic: title