sty1delta.h2o2.rna.2
Schizosaccharomyces pombe
Protocols: Cells were collected by filtration. Filters were immediately frozen in liquid nitrogen. Cells were grown for 15 min in 0.5 mM H2O2 Total RNA was extracted using a hot phenol extraction method as described in Lyne et al. (2003) BMC Genomics 4:27. Clean up and DNase treatment were performed on with Purelink RNA microcolumns (Life Technologies) following the manufacturer's instructions. Samples were ribo-depleted using Ribo-Zero Gold rRNA Removal Kit Yeast (Illumina) with a 4 ug input. 30 ng of ribo-depleted RNA was used as starting material for the NEXTflex Rapid Directional qRNA-Seq Kit (Bioo Scientific). Gel purified RNA fragments were treated with 10 units of T4 PNK (Thermo Fisher) in a low pH buffer (700 mM Tris pH 7, 50 mM DTT, 100 mM MgCl2) for 30 min at 37C. ATP and buffer A (Thermo Fisher) were then added for an additional 30 min incubation. RNA fragments were column-purified (PureLink RNA micro-columns, Life Technologies). 100 ng were used as input for the NEXTflex Small RNA Sequencing Kit v2 (Bioo Scientific), following manufacturer's protocol.
ENA-FIRST-PUBLIC
2020-06-01T04:03:53Z
ENA-LAST-UPDATE
2020-01-28T14:37:41Z
External Id
SAMEA6505906
INSDC center name
Department of Biochemistry University of Cambridge UK
INSDC first public
2020-06-01T04:03:53Z
INSDC last update
2020-01-28T14:37:41Z
INSDC status
public
Submitter Id
E-MTAB-8744:sty1delta.h2o2.rna.2
broker name
ArrayExpress
common name
fission yeast
fraction
rRNA depleted total RNA
genotype
sty1delta
replicate
2
sample name
E-MTAB-8744:sty1delta.h2o2.rna.2
scientific_name
Schizosaccharomyces pombe
stimulus
hydrogen peroxide