characteristics.text: 2016-04-30T17:09:59Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:21:17Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2189752
characteristics.characteristic: External Id
characteristics.text: EMBL-EBI
characteristics.characteristic: INSDC center name
characteristics.text: 2016-04-30T17:09:59Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:21:17Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS226993
characteristics.characteristic: SRA accession
characteristics.text: B730
characteristics.characteristic: StrainOrLine
characteristics.text: E-SYBR-6:B730
characteristics.characteristic: Submitter Id
characteristics.text: frozen samples
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000922
characteristics.characteristic: bio source type
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: baker's yeast
characteristics.characteristic: common name
characteristics.text: Protocols: Each sample was treated with one of the following control chemicals or sub-lethal drug doses: 1% DMSO (Dimethyl sulfoxide ), 200 uM 089 (a novel compound with fungicidal effects identified in the Stefanini lab), 2.5 uM ketoconazole or 50uM ketoconazole. After the 20th generation, cells were harvested and genomic DNAs were extracted using QIAamp DNA Mini Kit (Qiagen) following manufacturer’s instructions. Each 20-mer uptag barcode was amplified with an unique forward primer: UpTag U1 Forward. This primer has the U1 sequence plus the sequence R2 required for cluster formation on the Illumina flowcell. Twelve reverse primers were specifically designed with PerlPrimer v1.1.19 Software. The reverse composite primers contain sequences of the common barcode primers that will anneal upstream of U2 in a specific region of the kanamycin sequence, 12 different indexing tags of 6 nucleotides and the sequence R1 required for hybridization to the Illumina flowcell and for indexing tags reading. Each PCR amplification was carried out in 50 ul, using 100 ng of sample, 0.5uM of each forward and reverse primer (Oligosynthese BioTeZ, Germany) and 2.5 U of Invitrogen Accuprime Pfx DNA polymerase (Cat. No 12344-024) with the following cycling conditions: 1 cycle at 95 degree C for 3 min; and 25 amplification cycles (94 degree C for 30 s, 55 degree C for 30 s, 68 degree C for 30 s); followed by 68 degree C for 10 min. PCR products were then purified with Beckman Coulter SPRI beads (Cat. No. 082A63881) using a ratio of 1. After PCR purification, DNA was quantified with 7500 DNA chip on 2100 Bioanalyzer (Agilent). After amplification products were clustered in a single-end flow cell and sequenced on an Illumina GA2x - 36 bases for the read and 7 bases for the index. Equimolar amounts of PCR products were pooled.
characteristics.characteristic: description
characteristics.text: Saccharomyces cerevisiae
characteristics.characteristic: organism
characteristics.text: Saccharomyces cerevisiae
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_4932
characteristics.characteristic: organism
characteristics.text: Saccharomyces cerevisiae
characteristics.characteristic: scientific_name
characteristics.text: B730
characteristics.characteristic: title